Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Nucleotide and/or Amino Acid Sequence Disclosures
REQUIREMENTS FOR PATENT APPLICATIONS CONTAINING NUCLEOTIDE AND/OR AMINO ACID SEQUENCE DISCLOSURES
Items 1) and 2) provide general guidance related to requirements for sequence disclosures.
37 CFR 1.821(c) requires that patent applications which contain disclosures of nucleotide and/or amino acid sequences that fall within the definitions of 37 CFR 1.821(a) must contain a "Sequence Listing," as a separate part of the disclosure, which presents the nucleotide and/or amino acid sequences and associated information using the symbols and format in accordance with the requirements of 37 CFR 1.821 - 1.825. This "Sequence Listing" part of the disclosure may be submitted:
In accordance with 37 CFR 1.821(c)(1) via the USPTO patent electronic filing system (see Section I.1 of the Legal Framework for Patent Electronic System (https://www.uspto.gov/PatentLegalFramework), hereinafter "Legal Framework") as an ASCII text file, together with an incorporation-by-reference of the material in the ASCII text file in a separate paragraph of the specification as required by 37 CFR 1.823(b)(1) identifying:
the name of the ASCII text file;
ii) the date of creation; and
iii) the size of the ASCII text file in bytes;
In accordance with 37 CFR 1.821(c)(1) on read-only optical disc(s) as permitted by 37 CFR 1.52(e)(1)(ii), labeled according to 37 CFR 1.52(e)(5), with an incorporation-by-reference of the material in the ASCII text file according to 37 CFR 1.52(e)(8) and 37 CFR 1.823(b)(1) in a separate paragraph of the specification identifying:
the name of the ASCII text file;
the date of creation; and
the size of the ASCII text file in bytes;
In accordance with 37 CFR 1.821(c)(2) via the USPTO patent electronic filing system as a PDF file (not recommended); or
In accordance with 37 CFR 1.821(c)(3) on physical sheets of paper (not recommended).
When a “Sequence Listing” has been submitted as a PDF file as in 1(c) above (37 CFR 1.821(c)(2)) or on physical sheets of paper as in 1(d) above (37 CFR 1.821(c)(3)), 37 CFR 1.821(e)(1) requires a computer readable form (CRF) of the “Sequence Listing” in accordance with the requirements of 37 CFR 1.824.
If the "Sequence Listing" required by 37 CFR 1.821(c) is filed via the USPTO patent electronic filing system as a PDF, then 37 CFR 1.821(e)(1)(ii) or 1.821(e)(2)(ii) requires submission of a statement that the "Sequence Listing" content of the PDF copy and the CRF copy (the ASCII text file copy) are identical.
If the "Sequence Listing" required by 37 CFR 1.821(c) is filed on paper or read-only optical disc, then 37 CFR 1.821(e)(1)(ii) or 1.821(e)(2)(ii) requires submission of a statement that the "Sequence Listing" content of the paper or read-only optical disc copy and the CRF are identical.
Specific deficiencies and the required response to this Office Action are as follows:
Specific deficiency – Nucleotide and/or amino acid sequences appearing in the specification are not identified by sequence identifiers in accordance with 37 CFR 1.821(d). Specifically, no sequence identification has been provided for the amino acid sequence presented in line 2 of paragraph [0018] of the specification filed 09/21/2025.
Required response – Applicant must provide:
A substitute specification in compliance with 37 CFR 1.52, 1.121(b)(3) and 1.125 inserting the required sequence identifiers, consisting of:
A copy of the previously-submitted specification, with deletions shown with strikethrough or brackets and insertions shown with underlining (marked-up version);
A copy of the amended specification without markings (clean version); and
A statement that the substitute specification contains no new matter.
DETAILED ACTION
Status of Application/Election/Restrictions
Claims 1-3 are pending in this application and under examination in this office action.
Specification
The disclosure is objected to because of the following informalities: The recitation “The ferritin protein of SEQ ID NO:2….” is objected to because based on the sequence listing, the sequence of SEQ ID NO:2 is a nucleic acid sequence, not an amino acid sequence. Appropriate correction is required.
The attempt to incorporate subject matter into this application by reference to “the pRBASFer vector disclosed in ‘Choi, JW (2016) Secretion of ferritin protein….and its feed efficiency’…” in paragraph [0019] is ineffective because the root words “incorporate” and/or “reference” have been omitted, see 37 CFR 1.57(c)(1); the reference document is not clearly identified as required by 37 CFR 1.57(c)(2)).
The claims and specification depend on the description of “pRBASFer” and “pRBASFer-ori” to define a gene encoding a Periserrula leucophryna-derived ferritin protein encompassed by the claims, but since “pRBASFer” and “pRBASFer-ori” can be updated, the specification fails to define the claimed sequences and therefore the claims include any number of undefined variants. It is unclear which sequence, the initial or the updated, should be used in the claimed method. Thus, the claims are indefinite. Based on MPEP 608.01(p) Completeness, the recitation of “pRBASFer” and “pRBASFer-ori” to define a gene encoding a Periserrula leucophryna-derived ferritin protein in the claims is inappropriate because the genes/proteins as defined by “pRBASFer” and “pRBASFer-ori” are essential materials, which are required by the claimed method. Thus, the incorporation by reference as by recitation of “the pRBASFer vector disclosed in ‘Choi, JW (2016) Secretion of ferritin protein….and its feed efficiency’…” in paragraph [0019] is not effective.
Further, the specification depends on the description of GenBank accession numbers DQ207752 and ABA55730 (paragraph [0059] of the specification) to define the ferritin protein and nucleic acid encompassed by the claims, but since accession numbers can be updated, the specification fails to define the claimed sequences and therefore the claims include any number of undefined variants. For example, Accession No. X87832 has been updated eight times since its initial submission. It is unclear which sequence, the initial or the updated, should be used in the claimed method.
Based on MPEP 608.01(p) Completeness, the recitation of GenBank accession Nos. to define genes/proteins in the claims is inappropriate because the genes/proteins as defined by accession no. are essential materials, which are required by the claimed method. Thus, the incorporation by reference as by recitation of GenBank accession no. is not effective. The incorporation of essential material in the specification by reference to an unpublished U.S. application, foreign application or patent, or to a publication is improper. Applicant is required to amend the disclosure to include the material incorporated by reference, if the material is relied upon to overcome any objection, rejection, or other requirement imposed by the Office. The amendment must be accompanied by a statement executed by the applicant, or a practitioner representing the applicant, stating that the material being inserted is the material previously incorporated by reference and that the amendment contains no new matter. 37 CFR 1.57(f). Also See MPEP 608.01 (p). I. Incorporation by reference & 37CFR 1.57. Incorporation by reference:
“(c) “Essential material” may be incorporated by reference, but only by way of an incorporation by reference to a U.S. patent or U.S. patent application publication, which patent or patent application publication does not itself incorporate such essential material by reference. “Essential material” is material that is necessary to: (1) Provide a written description of the claimed invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and set forth the best mode contemplated by the inventor of carrying out the invention as required by the first paragraph of 35 U.S.C. 112; (2) Describe the claimed invention in terms that particularly point out and distinctly claim the invention as required by the second paragraph of 35 U.S.C. 112…”
The incorporation by reference will not be effective until correction is made to comply with 37 CFR 1.57(c), (d), or (e). If the incorporated material is relied upon to meet any outstanding objection, rejection, or other requirement imposed by the Office, the correction must be made within any time period set by the Office for responding to the objection, rejection, or other requirement for the incorporation to be effective. Compliance will not be held in abeyance with respect to responding to the objection, rejection, or other requirement for the incorporation to be effective. In no case may the correction be made later than the close of prosecution as defined in 37 CFR 1.114(b), or abandonment of the application, whichever occurs earlier.
Any correction inserting material by amendment that was previously incorporated by reference must be accompanied by a statement that the material being inserted is the material incorporated by reference and the amendment contains no new matter. 37 CFR 1.57(g).
Claim Objections
Claims 1 and 3 are objected to because of the following informalities: The recitations “PCR”, “T vector” “pRBASFer” “pRBASFer-ori” in claim 1 are not unique or common abbreviations in the art. Applicants are required to spell out “PCR” and “T vector” at the first usage. In addition, the symbol of “ferritin” gene should be italic. Appropriate correction is required.
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 1-3 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor, or for pre-AIA the applicant regards as the invention.
Claims 1-3 are indefinite because:
i. Regarding claim 1, the limitation “a gene encoding Periserrula leucophryna-derived ferritin protein represented by an amino acid sequence of SEQ ID NO:1…” recited in claim 1 is indefinite because it is unclear what other proteins and/or sequences are included and within the scope of the claim. Applicant fails to set forth the metes and bounds of what is encompassed within the limitation “a gene encoding.....represented by an amino acid sequence…”. Since the metes and bounds are unknown, a skilled artisan cannot envision what other proteins and/or sequences are included and within the scope of the claim.
The terms “T vector”, “pRBASFer” and “pRBASFer-ori” are recited in the claims without a reference to a precise amino acid sequence identified by a proper SEQ ID NO: or providing a full name for abbreviated names. Without identification of property or combination of properties which are unique to and, therefore, definitive of the instant recitations, the metes and bounds of the claims remain undetermined. Further, the use of laboratory designations only to identify a particular molecule renders the claims indefinite because different laboratories may use the same laboratory designations to define completely distinct molecules.
Further, the claims and specification depend on the description of “pRBASFer” and “pRBASFer-ori” to define a vector comprising a gene encoding a Periserrula leucophryna-derived ferritin protein encompassed by the claims, but since “pRBASFer” and “pRBASFer-ori” can be updated, the specification fails to define the claimed sequences and therefore the claims include any number of undefined variants. It is unclear which sequence, the initial or the updated, should be used in the claimed method. Thus, the claims are indefinite. The rejection can be obviated by amending the claims to specifically and uniquely identify “pRBASFer” and “pRBASFer-ori”, for example, by SEQ ID NO: and function of “pRBASFer” and “pRBASFer-ori”.
ii. Claim 1 recites the limitation "the replication origin" in line 5 of the claim. There is insufficient antecedent basis for this limitation in the claim.
iii. Claim 3 recites the limitation "the effect " in line 5 of the claim. There is insufficient antecedent basis for this limitation in the claim. In addition, The term “increased” in claim 3 is a relative term which renders the claim indefinite. The term “increased” is not defined by the claim, the specification does not provide a standard for ascertaining the requisite degree, and one of ordinary skill in the art would not be reasonably apprised of the scope of the invention. Applicant fails to set forth the metes and bounds of what is encompassed within the definition of “increased”. Since the metes and bounds are unknown, a skilled artisan cannot envision what would be considered as “a biological activity…. laid from laying hens are all increased”, what would be considered as “a biological activity…production index of broilers are increased” and what would be considered as “a biological activity…growth/feed (G/F) ratio of weanling pigs are increased” recited in the claim. Thus, the claim is indefinite.
iv. The rest of claims are indefinite as depending from an indefinite claim.
Claim Rejections - 35 USC § 112
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claims 1-3 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for pre-AIA the inventor(s), at the time the application was filed, had possession of the claimed invention.
To provide adequate written description and evidence of possession of a claimed genus, the specification must provide sufficient distinguishing identifying characteristics of the genus. The factors to be considered include disclosure of complete or partial structure, physical and/or chemical properties, functional characteristics, structure/function correlation, methods of making the claimed product, or any combination thereof.
Claims 1-3 encompass using a genus of restriction enzyme to digest the existing pRBASFer vector, and a genus of Periserrula leucophryna-derived ferritin protein represented by an amino acid sequence of SEQ ID NO:1, which encompasses any fragments within the amino acid sequence of SEQ ID NO:1 and other undefined sequences.
Applicant has not disclosed sufficient species for the broad genus of restriction enzyme to digest the existing pRBASFer vector to prepare the claimed new recombinant secretion vector: pRBASFer-ori or the broad genus of Periserrula leucophryna-derived protein represented by an amino acid sequence of SEQ ID NO:1.
The specification only describes: i) cloning the DNA fragment of the replication origin (SEQ ID NO: 2 encoding the Periserrula leucophryna-derived ferritin protein of SEQ ID NO:1) into a pGEM-Teasy vector (i.e. T-pRBori), wherein the DNA fragment of the replication origin was obtained by PCR using the claimed forward and reverse primers (SEQ ID NOs:5-6); ii) obtaining the DNA fragment of the replication origin by digesting the T-pRBori vector with EcoRV and PciI and iii) preparing a new recombinant secretion vector: pRBASFer-ori vector (i.e. comprising two replication origins encoding Periserrula leucophryna-derived ferritin protein) by digesting the existing pRBASFer vector with a restriction enzyme MluI and filling with a Klenow enzyme, and again digesting with another restriction enzyme, PciI and ligating the DNA fragment of the replication origin obtained from T-pRBori (see p. 13-14, Figure 2).
In making a determination of whether the application complies with the written description requirement of 35 U.S.C. 112, first paragraph, it is necessary to understand what Applicant is in possession of and what Applicant is claiming.
M.P.E.P. § 2163 instructs:
An invention described solely in terms of a method of making and/or its function may lack written descriptive support where there is no described or art-recognized correlation between the disclosed function and the structure(s) responsible for the function. . . .
An applicant may show possession of an invention by disclosure of drawings or structural chemical formulas that are sufficiently detailed to show that applicant was in possession of the claimed invention as a whole. . . .
An applicant may also show that an invention is complete by disclosure of sufficiently detailed, relevant identifying characteristics which provide evidence that applicant was in possession of the claimed invention, i.e., complete or partial structure, other physical and/or chemical properties, functional characteristics when coupled with a known or disclosed correlation between function and structure, or some combination of such characteristics.”
This standard has not been met in this case. From the specification, Applicant is in possession of using restriction enzymes EcoRV and PciI to obtain the DNA fragment of the replication origin (encoding the Periserrula leucophryna-derived ferritin protein of instant SEQ ID NO:1) from the T-pRBori vector and using a restriction enzyme MluI to digest the existing pRBASFer vector and filling with a Klenow enzyme, and again digesting with another restriction enzyme, PciI and ligating the DNA fragment of the replication origin obtained from T-pRBori to generate a new recombinant secretion vector: pRBASFer-ori vector (see p. 13-14, Figure 2).
However, Applicant is not in possession of using all kinds of restriction enzymes to prepare the new recombinant secretion vector: pRBASFer-ori vector because the DNA fragment of the replication origin from the T-pRBori vector was PCR using specific primers and obtained by digestion with specific restriction enzymes EcoRV and PciI to generate specific sites for ligation into the originally existing pRBASFer vector. The new recombinant secretion vector: pRBASFer-ori vector is generated by using a restriction enzyme MluI to digest the existing pRBASFer vector and filling with a Klenow enzyme, and again digesting with another restriction enzyme, PciI to generate restriction sites for ligating the DNA fragment of the replication origin obtained from the T-pRBori vector. Applicant is also not in possession of Periserrula leucophryna-derived ferritin protein comprising fragments within the amino acid sequence of SEQ ID NO:1 and other undefined sequences.
The specification provides no identification of any particular portion of the restriction sites and restriction enzymes that must be conserved to generate the claimed new recombinant secretion vector: pRBASFer-ori vector. The instant specification fails to provide sufficient descriptive information, such as definitive structural or functional features of the claimed genus of restriction enzymes for restriction sites to ligate the DNA fragment of the replication origin obtained from the T-pRBori vector which contains the PCR products using the specific forward and reverse primers of SEQ ID NOs: 5-6.
The specification provides no identification of any particular portion of the genus of Periserrula leucophryna-derived ferritin protein comprising fragments within the amino acid sequence of SEQ ID NO:1 and other undefined sequences that must be conserved. The instant specification fails to provide sufficient descriptive information, such as definitive structural or functional features of the claimed genus of Periserrula leucophryna-derived ferritin protein comprising fragments within the amino acid sequence of SEQ ID NO:1 and other undefined sequences.
There is no description of the conserved regions which are critical to the function of the claimed genus of restriction enzyme. There is no description of the sites at which variability may be tolerated and there is no information regarding the relation of structure of other restriction enzymes and restriction sites to the function of restriction enzymes EcoRV and PciI to obtain the DNA fragment of the replication origin from the T-pRBori vector and the function of restriction enzyme MluI and another restriction enzyme, PciI to digest the existing pRBASFer vector for ligating the DNA fragment of the replication origin from the T-pRBori vector.
There is no description of the conserved regions which are critical to the function of the claimed genus of Periserrula leucophryna-derived ferritin protein comprising fragments within the amino acid sequence of SEQ ID NO:1 and other undefined sequences. There is no description of the sites at which variability may be tolerated and there is no information regarding the relation of structure of other Periserrula leucophryna-derived ferritin protein comprising fragments within the amino acid sequence of SEQ ID NO:1 and other undefined sequences to the function of Periserrula leucophryna-derived ferritin protein comprising the amino acid sequence of SEQ ID NO:1.
Furthermore, the prior art does not provide compensatory structural or correlative teachings sufficient to enable one of skill to identify what other specific restriction enzymes for ligating the DNA fragment of the replication origin from the T-pRBori vector to the existing pRBASFer vector might be and what other Periserrula leucophryna-derived ferritin protein comprising fragments within the amino acid sequence of SEQ ID NO:1 and other undefined sequences might be. Since the common characteristics/features of other restriction enzymes and other Periserrula leucophryna-derived ferritin protein comprising fragments within the amino acid sequence of SEQ ID NO:1 and other undefined sequences are unknown, a skilled artisan cannot envision the functional correlations of the genus with the claimed invention. Accordingly, in the absence of sufficient recitation of distinguishing identifying characteristics, the specification does not provide adequate written description of the genus of restriction enzymes and the genus of Periserrula leucophryna-derived protein represented by an amino acid sequence of SEQ ID NO:1.
Based on MPEP § 2161.01 and §2163, “to satisfy the written description requirement, a patent specification must describe the claimed invention in sufficient detail that one skilled in the art can reasonably conclude that the inventor had possession of the claimed invention. See, e.g., Moba, B.V. v. Diamond Automation, Inc., 325 F.3d 1306, 1319, 66 USPQ2d 1429, 1438 (Fed. Cir. 2003); Vas-Cath, Inc. v. Mahurkar, 935 F.2d at 1563, 19 USPQ2d at 1116”.
Vas-Cath Inc. v. Mahurkar, 19USPQ2d 1111, clearly states “applicant must convey with reasonable clarity to those skilled in the art that, as of the filing date sought, he or she was in possession of the invention. The invention is, for purposes of the ‘written description’ inquiry, whatever is now claimed.” (See page 1117.) The specification does not “clearly allow persons of ordinary skill in the art to recognize that [he or she] invented what is claimed.” (See Vas-Cath at page 1116).
As discussed above, the skilled artisan cannot envision the detailed chemical structure of the encompassed genus of restriction enzymes and Periserrula leucophryna-derived protein represented by an amino acid sequence of SEQ ID NO:1, and therefore conception is not achieved until reduction to practice has occurred, regardless of the complexity or simplicity of the method of isolation. Adequate written description requires more than a mere statement that it is part of the invention and reference to a potential method of isolating it. The compound itself is required. See Fiers v. Revel, 25 USPQ2d 1601 at 1606 (CAFC 1993) and Amgen Inc. v. Chugai Pharmaceutical Co. Ltd., 18 USPQ2d 1016. One cannot describe what one has not conceived. See Fiddes v. Baird, 30 USPQ2d 1481 at 1483.
Therefore, the claimed method and composition have not met the written description provision of 35 U.S.C. §112, first paragraph. Applicant is reminded that Vas-Cath makes clear that the written description provision of 35 U.S.C. §112 is severable from its enablement provision (see page 1115). Applicant is directed to the Guidelines for the Examination of Patent Applications Under the 35 U.S.C. 112, ¶ 1 "Written Description" Requirement. See MPEP § 2161.01 and 2163.
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
Claim Rejections - 35 USC § 102
The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale or otherwise available to the public before the effective filing date of the claimed invention.
(a)(2) the claimed invention was described in a patent issued under section 151, or in an application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
Claims 1-3 are rejected under 35 U.S.C. 102(a)(1) & (a)(2) as being anticipated by Lee et al. (Korean Society for Biotechnology and Bioengineering Journal, 2019; 34: pp258-266; Citations are based on a Google translate-English version) as evidenced by Jeong et al. (J. Microbiol., 2006; 44 (1): 54-63).
Claims 1-3 are drawn to a mass production method of Periserrula leucophryna-derived ferritin protein using a recombinant vector comprising a gene encoding Periserrula leucophryna-derived ferritin protein having an amino acid sequence of SEQ ID NO: 1 and a composition comprising ferritin protein produced by the claimed method, and wherein the composition is a feed additive for laying hens, broilers and weanling pigs, and wherein the ferritin protein is added in amount of 0.01 to 5 parts by weight based on 100 parts by weight of main feed and drinking water (0.01-5%).
Lee et al. teach a mass production method of Periserrula leucophryna-derived ferritin protein using a recombinant vector, pRBSAFer-ori vector which comprises two replication origins encoding Periserrula leucophryna-derived ferritin protein having the amino acid sequence of instant SEQ ID NO: 1 (See p. 258, abstract; p.259, sections 2.2 to p. 260, section 2.7; p. 260-265, section: Results and discussion, Figure 1). Lee teaches that the mass production method comprises the steps recited in instant claim 1:
1) using a forward primer: 1-gggacatgttctttcctgcgttatcccctg- 30 (which is 100% identical to instant SEQ ID NO:5) and a reverse primer: 1-cccgatatcctatttagaatattgtttagt-30 (which is 100% identical to instant SEQ ID NO: 6) to PCR a DNA fragment of the replication origin from a pRBASFer vector that comprises a ferritin gene of Periserrula leucophryna, and cloning the DNA fragment of the replication origin into a pGEM-Teasy vector (i.e. T-pRBori) (see p. 259, section 2.3);
2) obtaining a DNA fragment of the replication origin by digesting the T-pRBori vector with EcoRV and PciI to prepare a replication origin fragment of 508bp; (p. 260, 1st col., 1st paragraph)
3) preparing a new recombinant secretion vector: pRBASFer-ori vector which with two replication origins, wherein the second replication origin obtained from step 2) is introduced in a tandem array by digesting the existing pRBASFer vector with a restriction enzyme MluI and filling with a Klenow enzyme, and again digesting with another restriction enzyme, PciI and ligating the DNA fragment of the second replication origin obtained in step 2) (see p.260-p.261,Figure 1); and introducing the pRBSAFer-ori vector into Bacillus subtilis LKS87 to generate a Bacillus subtilis transformant (see p. 260, 1st col., 1st paragraph); and
4) culturing the Bacillus subtilis transformant from step 3) in a PY liquid medium at 30oC at a stirring speed of 150rpm at an air injection rate of 1 vvm for72 hours wherein the PY liquid medium is supplemented with soy peptone 2% (w/v) as a nitrogen source and barley 2% (w/v) as a carbon source (see p.259, sections 2.2 to p. 260, section 2.7; p. 260-265, section: Results and discussion, Figure 1).
The pRBASFer vector disclosed by Lee comprises a ferritin gene having GenBank accession number DQ207752 (which is identical to GenBank accession number DQ207752 shown in paragraph [0059] of the instant specification) (see p.259, 1st col.), which encodes Periserrula leucophryna-derived protein having the amino acid sequence of instant SEQ ID NO:1 as evidenced by Jeong et al. (J. Microbiol., 2006; 44 (1): 54-63; see the sequence alignment below). Jeong teaches a method of cloning and expressing Ferritin gene of Periserrula leucophryna using a pT7-7 expression vector, wherein the Ferritin protein of Periserrula leucophryna has the amino acid sequence of instant SEQ ID NO:1 (see the sequence alignment below; p. 54, abstract; p. 55-56, Materials and Methods).
Lee also teaches a composition comprising the ferritin protein produced by the mass production method, and wherein the composition is a feed additive for laying hens, and wherein the ferritin protein is added in amount of 0.1% by weight of main feed and drinking water and wherein the effects of the composition as a feed additive for laying hens, broilers and weanling pigs is verified by increased eggshell color, freshness and iron content in eggshell and egg yolk measured by using Inductively Coupled Plasma emission spectroscopy (ICP) (see p.258, abstract; p. 263, section 3.3 to p. 265). Thus, claims 1-3 are anticipated by Lee.
The sequence search results disclose as follows:
SEQ ID NO:2
DQ207752
LOCUS DQ207752 1111 bp mRNA linear INV 15-MAR-2006
DEFINITION Periserrula leucophryna ferritin (PLF) mRNA, complete cds.
ACCESSION DQ207752
VERSION DQ207752.1
KEYWORDS .
SOURCE Periserrula leucophryna
ORGANISM Periserrula leucophryna
Eukaryota; Metazoa; Spiralia; Lophotrochozoa; Annelida; Polychaeta;
Errantia; Phyllodocida; Nereididae; Periserrula.
REFERENCE 1 (bases 1 to 1111)
AUTHORS Jeong,B.R., Chung,S.M., Baek,N.J., Koo,K.B., Baik,H.S., Joo,H.S.,
Chang,C.S. and Choi,J.W.
TITLE Characterization, cloning and expression of the ferritin gene from
the Korean polychaete, Periserrula leucophryna
JOURNAL J. Microbiol. 44 (1), 54-63 (2006)
PUBMED 16554718
REFERENCE 2 (bases 1 to 1111)
AUTHORS Koo,K.-B., Baek,N.-J., Paik,S.-Y., Yun,J.-W. and Choi,J.-W.
TITLE Direct Submission
JOURNAL Submitted (16-SEP-2005) Department of Bioindustry, College of Life
& Environment, Daegu University, 15 Naeri-Ri, Jinryang-up,
Kyungsan, Kyungbuk 712-714, South Korea
FEATURES Location/Qualifiers
source 1..1111
/organism="Periserrula leucophryna"
/mol_type="mRNA"
/isolate="Paik1977"
/db_xref="taxon:232278"
/geo_loc_name="South Korea: Yellow Sea"
/collection_date="1977"
/collected_by="E. I. Paik"
gene 1..1111
/gene="PLF"
CDS 129..653
/gene="PLF"
/note="iron binding protein"
/codon_start=1
/product="ferritin"
/protein_id="ABA55730.1"
/translation="MATSRQTMPRQNYHEECEAGINKQINLELYASYVYQSMAWYFNR
DDVALPGFHHFFKKASEEEREHAEKFMKYQNMRGGRIVLQDIKKPERDEWGTGLEAMQ
AAHALEKHVNQSLLDLHKLADGHDDGQLTDFLEGEYLKEQVEAIKEISDHITQLKRVG
PGLGEYMYDKELKS
Query Match 100.0%; Score 1111; Length 1111;
Best Local Similarity 100.0%;
Matches 1111; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 AAAACTTTGATATCGACTGTATCTTGGGACGTCAGTGTGCGTACGGATCGGCGGTATCTC 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 AAAACTTTGATATCGACTGTATCTTGGGACGTCAGTGTGCGTACGGATCGGCGGTATCTC 60
Qy 61 TTCTTCAAAACATCCTAACCATTTCTATTCAACGTCGTTAAGTCGAGTTCAATAGGTGCG 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 TTCTTCAAAACATCCTAACCATTTCTATTCAACGTCGTTAAGTCGAGTTCAATAGGTGCG 120
Qy 121 AACTAAAGATGGCCACATCCAGACAAACCATGCCCCGCCAGAACTACCATGAGGAGTGCG 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 AACTAAAGATGGCCACATCCAGACAAACCATGCCCCGCCAGAACTACCATGAGGAGTGCG 180
Qy 181 AAGCTGGAATCAACAAACAGATCAATCTCGAACTCTACGCCAGCTATGTTTACCAATCTA 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 AAGCTGGAATCAACAAACAGATCAATCTCGAACTCTACGCCAGCTATGTTTACCAATCTA 240
Qy 241 TGGCATGGTACTTCAACAGGGATGATGTTGCCCTCCCAGGCTTCCATCATTTCTTCAAGA 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 TGGCATGGTACTTCAACAGGGATGATGTTGCCCTCCCAGGCTTCCATCATTTCTTCAAGA 300
Qy 301 AGGCTTCTGAGGAAGAACGCGAACATGCTGAGAAGTTCATGAAGTACCAGAACATGAGGG 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 AGGCTTCTGAGGAAGAACGCGAACATGCTGAGAAGTTCATGAAGTACCAGAACATGAGGG 360
Qy 361 GTGGTCGTATCGTTCTGCAGGACATCAAGAAGCCGGAGAGGGATGAGTGGGGAACTGGAT 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 GTGGTCGTATCGTTCTGCAGGACATCAAGAAGCCGGAGAGGGATGAGTGGGGAACTGGAT 420
Qy 421 TGGAGGCCATGCAAGCGGCCCATGCACTGGAGAAGCATGTCAACCAGTCCCTGCTTGATC 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 TGGAGGCCATGCAAGCGGCCCATGCACTGGAGAAGCATGTCAACCAGTCCCTGCTTGATC 480
Qy 481 TTCACAAGTTGGCTGATGGCCACGATGACGGCCAGCTGACTGACTTCCTGGAGGGCGAGT 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 TTCACAAGTTGGCTGATGGCCACGATGACGGCCAGCTGACTGACTTCCTGGAGGGCGAGT 540
Qy 541 ACCTCAAGGAACAAGTAGAGGCAATCAAGGAGATCAGCGACCACATCACCCAGCTGAAAC 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 ACCTCAAGGAACAAGTAGAGGCAATCAAGGAGATCAGCGACCACATCACCCAGCTGAAAC 600
Qy 601 GTGTCGGTCCCGGCCTGGGAGAGTACATGTACGACAAGGAACTCAAGAGCTAGATGACCT 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 601 GTGTCGGTCCCGGCCTGGGAGAGTACATGTACGACAAGGAACTCAAGAGCTAGATGACCT 660
Qy 661 ACCTATAAGGTCAAGAGCCTCAGGCCGTCACCGAGACGCCAAGCATAGACCCACAATACT 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 661 ACCTATAAGGTCAAGAGCCTCAGGCCGTCACCGAGACGCCAAGCATAGACCCACAATACT 720
Qy 721 CTCGCTGCAGTCTTATCTCCTTAGCCTACCCCTGTATGAATCAACATCTTTGTTTGCTAT 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 721 CTCGCTGCAGTCTTATCTCCTTAGCCTACCCCTGTATGAATCAACATCTTTGTTTGCTAT 780
Qy 781 AGAATAGTCATCAGTAACAACATTCTCTTAATATCTATGATTTCTGCTTAGTGTGGTTCA 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 781 AGAATAGTCATCAGTAACAACATTCTCTTAATATCTATGATTTCTGCTTAGTGTGGTTCA 840
Qy 841 AGTTTAATACTTCTTAAAAGTTACACATGGCACGGCATGTGAAGATAATGGCATCTGTAG 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 841 AGTTTAATACTTCTTAAAAGTTACACATGGCACGGCATGTGAAGATAATGGCATCTGTAG 900
Qy 901 TTTCACATATTGCTTCTGAACTGTACCATAGTCAGGAAACTTAATATAACATTCCTAATG 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 901 TTTCACATATTGCTTCTGAACTGTACCATAGTCAGGAAACTTAATATAACATTCCTAATG 960
Qy 961 TCAAATCCAGGTACATTTATCAAATTTGTTTCAATCTTGTGTAACATTTGAAACTGCCCT 1020
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 961 TCAAATCCAGGTACATTTATCAAATTTGTTTCAATCTTGTGTAACATTTGAAACTGCCCT 1020
Qy 1021 GGTGGTAGAAGTTTGAAAAGAAAGGCCTTGGTATTGTTCAGTTGTGTTGAATAGATGCCT 1080
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1021 GGTGGTAGAAGTTTGAAAAGAAAGGCCTTGGTATTGTTCAGTTGTGTTGAATAGATGCCT 1080
Qy 1081 AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA 1111
|||||||||||||||||||||||||||||||
Db 1081 AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA 1111
SEQ ID NO:1
Q3HM65_PARUS
ID Q3HM65_PARUS Unreviewed; 174 AA.
AC Q3HM65;
DT 08-NOV-2005, integrated into UniProtKB/TrEMBL.
DT 08-NOV-2005, sequence version 1.
DT 02-APR-2025, entry version 71.
DE RecName: Full=Ferritin {ECO:0000256|RuleBase:RU361145};
DE EC=1.16.3.1 {ECO:0000256|RuleBase:RU361145};
GN Name=PLF {ECO:0000313|EMBL:ABA55730.1};
OS Paraleonnates uschakovi (Giant mud worm) (Periserrula leucophryna).
OC Eukaryota; Metazoa; Spiralia; Lophotrochozoa; Annelida; Polychaeta;
OC Errantia; Phyllodocida; Nereididae; Periserrula.
OX NCBI_TaxID=232278 {ECO:0000313|EMBL:ABA55730.1};
RN [1] {ECO:0000313|EMBL:ABA55730.1}
RP NUCLEOTIDE SEQUENCE.
RC STRAIN=Paik1977 {ECO:0000313|EMBL:ABA55730.1};
RA Koo K.-B., Baek N.-J., Paik S.-Y., Yun J.-W., Choi J.-W.;
RL Submitted (SEP-2005) to the EMBL/GenBank/DDBJ databases.
RN [2] {ECO:0000313|EMBL:ABA55730.1}
RP NUCLEOTIDE SEQUENCE.
RC STRAIN=Paik1977 {ECO:0000313|EMBL:ABA55730.1};
RX PubMed=16554718;
RA Jeong B.R., Chung S.M., Baek N.J., Koo K.B., Baik H.S., Joo H.S.,
RA Chang C.S., Choi J.W.;
RT "Characterization, cloning and expression of the ferritin gene from the
RT Korean polychaete, Periserrula leucophryna.";
RL J. Microbiol. 44:54-63(2006).
CC -!- FUNCTION: Stores iron in a soluble, non-toxic, readily available form.
CC Important for iron homeostasis. Has ferroxidase activity. Iron is taken
CC up in the ferrous form and deposited as ferric hydroxides after
CC oxidation. {ECO:0000256|ARBA:ARBA00025111}.
CC -!- FUNCTION: Stores iron in a soluble, non-toxic, readily available form.
CC Important for iron homeostasis. Iron is taken up in the ferrous form
CC and deposited as ferric hydroxides after oxidation.
CC {ECO:0000256|RuleBase:RU361145}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=4 Fe(2+) + O2 + 4 H(+) = 4 Fe(3+) + 2 H2O;
CC Xref=Rhea:RHEA:11148, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:29033, ChEBI:CHEBI:29034; EC=1.16.3.1;
CC Evidence={ECO:0000256|ARBA:ARBA00047990,
CC ECO:0000256|RuleBase:RU361145};
CC -!- SIMILARITY: Belongs to the ferritin family.
CC {ECO:0000256|ARBA:ARBA00007513, ECO:0000256|RuleBase:RU361145}.
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; DQ207752; ABA55730.1; -; mRNA.
DR AlphaFoldDB; Q3HM65; -.
DR SMR; Q3HM65; -.
DR GO; GO:0005737; C:cytoplasm; IEA:TreeGrafter.
DR GO; GO:0008199; F:ferric iron binding; IEA:InterPro.
DR GO; GO:0008198; F:ferrous iron binding; IEA:TreeGrafter.
DR GO; GO:0004322; F:ferroxidase activity; IEA:UniProtKB-EC.
DR GO; GO:0006879; P:intracellular iron ion homeostasis; IEA:UniProtKB-KW.
DR GO; GO:0006826; P:iron ion transport; IEA:InterPro.
DR CDD; cd01056; Euk_Ferritin; 1.
DR FunFam; 1.20.1260.10:FF:000002; Ferritin, mitochondrial; 1.
DR Gene3D; 1.20.1260.10; -; 1.
DR InterPro; IPR001519; Ferritin.
DR InterPro; IPR012347; Ferritin-like.
DR InterPro; IPR009040; Ferritin-like_diiron.
DR InterPro; IPR009078; Ferritin-like_SF.
DR InterPro; IPR014034; Ferritin_CS.
DR InterPro; IPR008331; Ferritin_DPS_dom.
DR PANTHER; PTHR11431; FERRITIN; 1.
DR PANTHER; PTHR11431:SF75; FERRITIN; 1.
DR Pfam; PF00210; Ferritin; 1.
DR SUPFAM; SSF47240; Ferritin-like; 1.
DR PROSITE; PS00540; FERRITIN_1; 1.
DR PROSITE; PS00204; FERRITIN_2; 1.
DR PROSITE; PS50905; FERRITIN_LIKE; 1.
PE 2: Evidence at transcript level;
KW Iron {ECO:0000256|ARBA:ARBA00023004, ECO:0000256|RuleBase:RU361145};
KW Iron storage {ECO:0000256|ARBA:ARBA00022434,
KW ECO:0000256|RuleBase:RU361145};
KW Metal-binding {ECO:0000256|ARBA:ARBA00022723,
KW ECO:0000256|RuleBase:RU361145};
KW Oxidoreductase {ECO:0000256|RuleBase:RU361145}.
FT DOMAIN 11..160
FT /note="Ferritin-like diiron"
FT /evidence="ECO:0000259|PROSITE:PS50905"
SQ SEQUENCE 174 AA; 20294 MW; 6388230E26661FFD CRC64;
Query Match 100.0%; Score 925; Length 174;
Best Local Similarity 100.0%;
Matches 174; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MATSRQTMPRQNYHEECEAGINKQINLELYASYVYQSMAWYFNRDDVALPGFHHFFKKAS 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MATSRQTMPRQNYHEECEAGINKQINLELYASYVYQSMAWYFNRDDVALPGFHHFFKKAS 60
Qy 61 EEEREHAEKFMKYQNMRGGRIVLQDIKKPERDEWGTGLEAMQAAHALEKHVNQSLLDLHK 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 EEEREHAEKFMKYQNMRGGRIVLQDIKKPERDEWGTGLEAMQAAHALEKHVNQSLLDLHK 120
Qy 121 LADGHDDGQLTDFLEGEYLKEQVEAIKEISDHITQLKRVGPGLGEYMYDKELKS 174
||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 LADGHDDGQLTDFLEGEYLKEQVEAIKEISDHITQLKRVGPGLGEYMYDKELKS 174
Claims 1-3 are rejected under 35 U.S.C. 102(a)(1) & (a)(2) as being anticipated by Choi (KR-102346499, published Jan 3, 2022, priority Aug 26, 2020, citations are based on KR-102346499-English translated version).
Choi (KR-102346499) teaches a method of mass production of Periserrula leucophryna-derived ferritin protein using a recombinant vector comprising a gene encoding Periserrula leucophryna-derived ferritin protein having an amino acid sequence of instant SEQ ID NO: 1 (see the sequence alignment below), and a composition comprising ferritin protein produced by the claimed method, and wherein the composition is a feed additive for laying hens, broilers and weanling pigs, and wherein the ferritin protein is added in amount of 0.01 to 5 parts by weight based on 100 parts by weight of main feed and drinking water (0.01-5%) (p.16-17, [0202]-[0208]). Choi teaches that the mass production method comprises steps of: 1) PCR a DNA fragment of the replication origin from a pRBASFer vector that comprises a ferritin gene of Periserrula leucophryna (encoding instant SEQ ID NO:2) by using a forward primer of instant SEQ ID NO: 5 and a reverse primer of instant SEQ ID NO: 6 (see the sequence alignment below), and cloning the DNA fragment of the replication origin into a pGEM-Teasy vector (i.e. T-pRBori); 2) obtaining the DNA fragment of the replication origin by digesting the T-pRBori vector with EcoRV and PciI to prepare a replication origin fragment of 508bp; 3) preparing a new recombinant secretion vector: pRBASFer-ori vector which comprises two replication origins wherein the second replication origin is introduced in a tandem array by digesting the existing pRBASFer vector with a restriction enzyme MluI and filling with a Klenow enzyme, and again digesting with another restriction enzyme, PciI and ligating the DNA fragment of the replication orign obtained in step 2); and introducing the pRBSAFer-ori vector into Bacillus subtilis LKS87 to generate a Bacillus subtilis transformant; and 4) culturing the Bacillus subtilis transformant from step 3) in a PY liquid medium at 30oC or 28oC-35oC at a stirring speed of 200rpm or 150-250rpm 200 rpm, at an air injection rate of 1 vvm or 0.5-2 vvm for 48 hours or 36-96 hours wherein the PY liquid medium is supplemented with soy peptone 1% to 4% (w/v) as a nitrogen source and barley 1% to 3% (w/v) as a carbon source (see p. 3, claims 3, 6; p.4-6, paragraphs [0001]; [0024]-[0065]; p. 8-17, [0080]-[208], drawing 2).
Choi also teaches a composition comprising the ferritin protein produced by the mass production method, and wherein the composition is a feed additive for laying hens, broilers and weanling pigs, and wherein the ferritin protein is added in amount of 0.01 to 5 parts by weight based on 100 parts by weight of main feed and drinking water (0.01-5%) and wherein the effects of the composition as a feed additive for laying hens, broilers and weanling pigs is verified by increased eggshell color, freshness and iron content in eggshell and egg yolk measured by using Inductively Coupled Plasma (ICP) and the change overtime after administration of ferritin-containing drinking water for 6 weeks compared to before the administration of ferritin-containing drinking water; for broilers is verified by increased survival rate, feed efficiency and production index of broilers, and for weanling pigs is verified average daily gain and growth/feed (G/F) ratio (see p. 12, [0158]-p. 17, [0208]). Thus, claims 1-3 are anticipated by Choi (KR-102346499).
The sequence search results disclose as follows:
SEQ ID NO:1
BKP52481
(NOTE: this sequence has 1 duplicate in the database searched)
ID BKP52481 standard; protein; 174 AA.
XX
AC BKP52481;
XX
DT 30-JUN-2022 (revised)
DT 17-MAR-2022 (first entry)
XX
DE Periserrula leucophryna ferritin protein, SEQ 1.
XX
KW Ferritin; feed-additive; protein production.
XX
OS Periserrula leucophryna.
XX
CC PN KR2346499-B1.
XX
CC PD 03-JAN-2022.
XX
CC PF 26-AUG-2020; 2020KR-00107888.
XX
PR 26-AUG-2020; 2020KR-00107888.
XX
CC PA (TURT-) TURTLEBIO CO LTD.
XX
CC PI Choi J;
XX
DR WPI; 2022-016972/008.
DR N-PSDB; BKP52482, BKP52483.
XX
CC PT Performing mass production of ferritin protein derived from Periserrula
CC PT leucophryna used in feed additive composition, by using recombinant
CC PT vector, cloning DNA fragment, obtaining DNA fragment from T vector,
CC PT cutting fragment, filling with Klenow enzyme, and culturing Bacillus
CC PT subtilis transformant.
XX
CC PS Claim 3; SEQ ID NO 1; 24pp; Korean.
XX
CC The present invention relates to a method for performing mass production
CC of ferritin protein of SEQ ID NO:1 (see BKP52481) derived from
CC Periserrula leucophryna. The invention also provides a composition
CC comprising the ferritin protein for feed addition for laying hens. The
CC method of the invention prepares a composition with excellent biological
CC activity, preferably superior biological activity in laying hens.
CC
CC Revised record issued on 30-JUN-2022 : Addition of DWPI-enhanced title
CC (PT field).
XX
SQ Sequence 174 AA;
Query Match 100.0%; Score 925; Length 174;
Best Local Similarity 100.0%;
Matches 174; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MATSRQTMPRQNYHEECEAGINKQINLELYASYVYQSMAWYFNRDDVALPGFHHFFKKAS 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MATSRQTMPRQNYHEECEAGINKQINLELYASYVYQSMAWYFNRDDVALPGFHHFFKKAS 60
Qy 61 EEEREHAEKFMKYQNMRGGRIVLQDIKKPERDEWGTGLEAMQAAHALEKHVNQSLLDLHK 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 EEEREHAEKFMKYQNMRGGRIVLQDIKKPERDEWGTGLEAMQAAHALEKHVNQSLLDLHK 120
Qy 121 LADGHDDGQLTDFLEGEYLKEQVEAIKEISDHITQLKRVGPGLGEYMYDKELKS 174
||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 LADGHDDGQLTDFLEGEYLKEQVEAIKEISDHITQLKRVGPGLGEYMYDKELKS 174
SEQ ID NO:2
BKP52482
(NOTE: this sequence has 1 duplicate in the database searched)
ID BKP52482 standard; DNA; 1111 BP.
XX
AC BKP52482;
XX
DT 30-JUN-2022 (revised)
DT 17-MAR-2022 (first entry)
XX
DE Periserrula leucophryna ferritin protein encoding DNA, SEQ 2.
XX
KW Ferritin gene; ds; feed-additive; gene; protein production.
XX
OS Periserrula leucophryna.
XX
FH Key Location/Qualifiers
FT CDS 129..653
FT /*tag= a
FT /product= "Ferritin"
XX
CC PN KR2346499-B1.
XX
CC PD 03-JAN-2022.
XX
CC PF 26-AUG-2020; 2020KR-00107888.
XX
PR 26-AUG-2020; 2020KR-00107888.
XX
CC PA (TURT-) TURTLEBIO CO LTD.
XX
CC PI Choi J;
XX
DR WPI; 2022-016972/008.
DR P-PSDB; BKP52481.
XX
CC PT Performing mass production of ferritin protein derived from Periserrula
CC PT leucophryna used in feed additive composition, by using recombinant
CC PT vector, cloning DNA fragment, obtaining DNA fragment from T vector,
CC PT cutting fragment, filling with Klenow enzyme, and culturing Bacillus
CC PT subtilis transformant.
XX
CC PS Disclosure; SEQ ID NO 2; 24pp; Korean.
XX
CC The present invention relates to a method for performing mass production
CC of ferritin protein of SEQ ID NO:1 (see BKP52481) derived from
CC Periserrula leucophryna. The invention also provides a composition
CC comprising the ferritin protein for feed addition for laying hens. The
CC method of the invention prepares a composition with excellent biological
CC activity, preferably superior biological activity in laying hens.
CC
CC Revised record issued on 30-JUN-2022 : Addition of DWPI-enhanced title
CC (PT field).
XX
SQ Sequence 1111 BP; 342 A; 246 C; 247 G; 276 T; 0 U; 0 Other;
Query Match 100.0%; Score 1111; Length 1111;
Best Local Similarity 100.0%;
Matches 1111; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 AAAACTTTGATATCGACTGTATCTTGGGACGTCAGTGTGCGTACGGATCGGCGGTATCTC 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 AAAACTTTGATATCGACTGTATCTTGGGACGTCAGTGTGCGTACGGATCGGCGGTATCTC 60
Qy 61 TTCTTCAAAACATCCTAACCATTTCTATTCAACGTCGTTAAGTCGAGTTCAATAGGTGCG 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 TTCTTCAAAACATCCTAACCATTTCTATTCAACGTCGTTAAGTCGAGTTCAATAGGTGCG 120
Qy 121 AACTAAAGATGGCCACATCCAGACAAACCATGCCCCGCCAGAACTACCATGAGGAGTGCG 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 AACTAAAGATGGCCACATCCAGACAAACCATGCCCCGCCAGAACTACCATGAGGAGTGCG 180
Qy 181 AAGCTGGAATCAACAAACAGATCAATCTCGAACTCTACGCCAGCTATGTTTACCAATCTA 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 AAGCTGGAATCAACAAACAGATCAATCTCGAACTCTACGCCAGCTATGTTTACCAATCTA 240
Qy 241 TGGCATGGTACTTCAACAGGGATGATGTTGCCCTCCCAGGCTTCCATCATTTCTTCAAGA 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 TGGCATGGTACTTCAACAGGGATGATGTTGCCCTCCCAGGCTTCCATCATTTCTTCAAGA 300
Qy 301 AGGCTTCTGAGGAAGAACGCGAACATGCTGAGAAGTTCATGAAGTACCAGAACATGAGGG 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 AGGCTTCTGAGGAAGAACGCGAACATGCTGAGAAGTTCATGAAGTACCAGAACATGAGGG 360
Qy 361 GTGGTCGTATCGTTCTGCAGGACATCAAGAAGCCGGAGAGGGATGAGTGGGGAACTGGAT 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 GTGGTCGTATCGTTCTGCAGGACATCAAGAAGCCGGAGAGGGATGAGTGGGGAACTGGAT 420
Qy 421 TGGAGGCCATGCAAGCGGCCCATGCACTGGAGAAGCATGTCAACCAGTCCCTGCTTGATC 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 TGGAGGCCATGCAAGCGGCCCATGCACTGGAGAAGCATGTCAACCAGTCCCTGCTTGATC 480
Qy 481 TTCACAAGTTGGCTGATGGCCACGATGACGGCCAGCTGACTGACTTCCTGGAGGGCGAGT 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 TTCACAAGTTGGCTGATGGCCACGATGACGGCCAGCTGACTGACTTCCTGGAGGGCGAGT 540
Qy 541 ACCTCAAGGAACAAGTAGAGGCAATCAAGGAGATCAGCGACCACATCACCCAGCTGAAAC 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 ACCTCAAGGAACAAGTAGAGGCAATCAAGGAGATCAGCGACCACATCACCCAGCTGAAAC 600
Qy 601 GTGTCGGTCCCGGCCTGGGAGAGTACATGTACGACAAGGAACTCAAGAGCTAGATGACCT 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 601 GTGTCGGTCCCGGCCTGGGAGAGTACATGTACGACAAGGAACTCAAGAGCTAGATGACCT 660
Qy 661 ACCTATAAGGTCAAGAGCCTCAGGCCGTCACCGAGACGCCAAGCATAGACCCACAATACT 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 661 ACCTATAAGGTCAAGAGCCTCAGGCCGTCACCGAGACGCCAAGCATAGACCCACAATACT 720
Qy 721 CTCGCTGCAGTCTTATCTCCTTAGCCTACCCCTGTATGAATCAACATCTTTGTTTGCTAT 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 721 CTCGCTGCAGTCTTATCTCCTTAGCCTACCCCTGTATGAATCAACATCTTTGTTTGCTAT 780
Qy 781 AGAATAGTCATCAGTAACAACATTCTCTTAATATCTATGATTTCTGCTTAGTGTGGTTCA 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 781 AGAATAGTCATCAGTAACAACATTCTCTTAATATCTATGATTTCTGCTTAGTGTGGTTCA 840
Qy 841 AGTTTAATACTTCTTAAAAGTTACACATGGCACGGCATGTGAAGATAATGGCATCTGTAG 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 841 AGTTTAATACTTCTTAAAAGTTACACATGGCACGGCATGTGAAGATAATGGCATCTGTAG 900
Qy 901 TTTCACATATTGCTTCTGAACTGTACCATAGTCAGGAAACTTAATATAACATTCCTAATG 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 901 TTTCACATATTGCTTCTGAACTGTACCATAGTCAGGAAACTTAATATAACATTCCTAATG 960
Qy 961 TCAAATCCAGGTACATTTATCAAATTTGTTTCAATCTTGTGTAACATTTGAAACTGCCCT 1020
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 961 TCAAATCCAGGTACATTTATCAAATTTGTTTCAATCTTGTGTAACATTTGAAACTGCCCT 1020
Qy 1021 GGTGGTAGAAGTTTGAAAAGAAAGGCCTTGGTATTGTTCAGTTGTGTTGAATAGATGCCT 1080
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1021 GGTGGTAGAAGTTTGAAAAGAAAGGCCTTGGTATTGTTCAGTTGTGTTGAATAGATGCCT 1080
Qy 1081 AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA 1111
|||||||||||||||||||||||||||||||
Db 1081 AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA 1111
SEQ ID NO:5
BKP52485
(NOTE: this sequence has 1 duplicate in the database searched)
ID BKP52485 standard; DNA; 30 BP.
XX
AC BKP52485;
XX
DT 30-JUN-2022 (revised)
DT 17-MAR-2022 (first entry)
XX
DE Replication origin forward PCR primer, SEQ 5.
XX
KW PCR; feed-additive; primer; protein production; ss.
XX
OS Unidentified.
XX
CC PN KR2346499-B1.
XX
CC PD 03-JAN-2022.
XX
CC PF 26-AUG-2020; 2020KR-00107888.
XX
PR 26-AUG-2020; 2020KR-00107888.
XX
CC PA (TURT-) TURTLEBIO CO LTD.
XX
CC PI Choi J;
XX
DR WPI; 2022-016972/008.
XX
CC PT Performing mass production of ferritin protein derived from Periserrula
CC PT leucophryna used in feed additive composition, by using recombinant
CC PT vector, cloning DNA fragment, obtaining DNA fragment from T vector,
CC PT cutting fragment, filling with Klenow enzyme, and culturing Bacillus
CC PT subtilis transformant.
XX
CC PS Claim 3; SEQ ID NO 5; 24pp; Korean.
XX
CC The present invention relates to a method for performing mass production
CC of ferritin protein of SEQ ID NO:1 (see BKP52481) derived from
CC Periserrula leucophryna. The invention also provides a composition
CC comprising the ferritin protein for feed addition for laying hens. The
CC method of the invention prepares a composition with excellent biological
CC activity, preferably superior biological activity in laying hens.
CC
CC Revised record issued on 30-JUN-2022 : Addition of DWPI-enhanced title
CC (PT field).
XX
SQ Sequence 30 BP; 3 A; 9 C; 7 G; 11 T; 0 U; 0 Other;
Query Match 100.0%; Score 30; Length 30;
Score over Length 100.0%;
Best Local Similarity 100.0%;
Matches 30; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 GGGACATGTTCTTTCCTGCGTTATCCCCTG 30
||||||||||||||||||||||||||||||
Db 1 GGGACATGTTCTTTCCTGCGTTATCCCCTG 30
SEQ ID NO:6
BKP52486
(NOTE: this sequence has 1 duplicate in the database searched)
ID BKP52486 standard; DNA; 30 BP.
XX
AC BKP52486;
XX
DT 30-JUN-2022 (revised)
DT 17-MAR-2022 (first entry)
XX
DE Replication origin reverse PCR primer, SEQ 6.
XX
KW PCR; feed-additive; primer; protein production; ss.
XX
OS Unidentified.
XX
CC PN KR2346499-B1.
XX
CC PD 03-JAN-2022.
XX
CC PF 26-AUG-2020; 2020KR-00107888.
XX
PR 26-AUG-2020; 2020KR-00107888.
XX
CC PA (TURT-) TURTLEBIO CO LTD.
XX
CC PI Choi J;
XX
DR WPI; 2022-016972/008.
XX
CC PT Performing mass production of ferritin protein derived from Periserrula
CC PT leucophryna used in feed additive composition, by using recombinant
CC PT vector, cloning DNA fragment, obtaining DNA fragment from T vector,
CC PT cutting fragment, filling with Klenow enzyme, and culturing Bacillus
CC PT subtilis transformant.
XX
CC PS Claim 3; SEQ ID NO 6; 24pp; Korean.
XX
CC The present invention relates to a method for performing mass production
CC of ferritin protein of SEQ ID NO:1 (see BKP52481) derived from
CC Periserrula leucophryna. The invention also provides a composition
CC comprising the ferritin protein for feed addition for laying hens. The
CC method of the invention prepares a composition with excellent biological
CC activity, preferably superior biological activity in laying hens.
CC
CC Revised record issued on 30-JUN-2022 : Addition of DWPI-enhanced title
CC (PT field).
XX
SQ Sequence 30 BP; 8 A; 5 C; 4 G; 13 T; 0 U; 0 Other;
Query Match 100.0%; Score 30; Length 30;
Score over Length 100.0%;
Best Local Similarity 100.0%;
Matches 30; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 CCCGATATCCTATTTAGAATATTGTTTAGT 30
||||||||||||||||||||||||||||||
Db 1 CCCGATATCCTATTTAGAATATTGTTTAGT 30
Conclusion
NO CLAIM IS ALLOWED.
The prior art made of record and not relied upon is considered pertinent to applicant's disclosure.
Choi (Korean Society for Biotechnology and Bioengineering Journal 31(2): 105-112 (2016). dx.doi.org/10.7841/ksbbj.2016.31.2.105; ISSN 1225-7117 / eISSN 2288-8268) teaches a method of generating a recombinant pRBAS-PLF vector and a Bacillus subtilis LKS strain transformant harboring the pRBAS-PLF vector to produce Periserrula leucophryna-derived ferritin protein and culturing the Bacillus subtilis LKS strain transformant in a PY medium supplemented with 1% peptone, 0.5% yeastex LSP, 2% Monodex (see p.105, abstract; p.106-107, Materials and Methods).
Kim et al. (Mol. Cells, 1997; 7:158-164) teach a heterologous protein secretion system of Bacillus subtilis (see p. 158, abstract; p. 159-160, Materials and Methods).
Opatowsky et al. (US/15258425) teach an oligonucleotide primer set comprising a forward primer having the sequence of SEQ Id NO:39, which is 94.7% identical to instant SEQ ID NO:5 (see the sequence alignment below).
SEQ ID NO:5
US-15-258-425A-39
Sequence 39, US/15258425A
GENERAL INFORMATION
APPLICANT: Bar Ilan University
APPLICANT: Opatowsky, Yarden
APPLICANT: Yerushalmi, Gal
APPLICANT: Blus-Kadosh, Inna
APPLICANT: Neznansky, Avi
APPLICANT: Banin, Ehud
TITLE OF INVENTION: ANTIMICORBIAL COMPOSITIONS AND METHODS AND USES THEREOF
FILE REFERENCE: 2433411
CURRENT APPLICATION NUMBER: US/15/258,425A
CURRENT FILING DATE: 2016-09-07
PRIOR APPLICATION NUMBER: US 62/215,196
PRIOR FILING DATE: 2015-09-08
NUMBER OF SEQ ID NOS: 56
SEQ ID NO 39
LENGTH: 31
TYPE: DNA
ORGANISM: Artificial Sequence
FEATURE:
OTHER INFORMATION: Primer 1325
Query Match 94.7%; Score 28.4; Length 31;
Score over Length 91.6%;
Best Local Similarity 96.7%;
Matches 29; Conservative 0; Mismatches 1; Indels 0; Gaps 0;
Qy 1 GGGACATGTTCTTTCCTGCGTTATCCCCTG 30
| ||||||||||||||||||||||||||||
Db 2 GAGACATGTTCTTTCCTGCGTTATCCCCTG 31
Any inquiry concerning this communication or earlier communications from the examiner should be directed to CHANG-YU WANG whose telephone number is (571)272-4521. The examiner can normally be reached on Monday-Thursday, 7:00am-5:00pm EST.
Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Jeffrey Stucker can be reached on 571-272-0911. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
Information regarding the status of an application may be obtained from the Patent Application Information Retrieval (PAIR) system. Status information for published applications may be obtained from either Private PAIR or Public PAIR. Status information for unpublished applications is available through Private PAIR only. For more information about the PAIR system, see https://ppair-my.uspto.gov/pair/PrivatePair. Should you have questions on access to the Private PAIR system, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative or access to the automated information system, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000.
Chang-Yu Wang
March 20, 2026
/CHANG-YU WANG/Primary Examiner, Art Unit 1675