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 .
DETAILED ACTION
This application is 371 of PCT/EP2022/081589.
The amendment filed on January 28, 2025 has been entered.
Status of Claims
Claims 1-20 are pending.
Claims 1-20 are under examination.
Foreign Priority
Receipt is acknowledged of certified copies of papers required by 37 CFR 1.55.
Information Disclosure Statement
The information disclosure statement (IDS) submitted on May 10, 2024 is in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement is being considered by the examiner.
Nucleotide and/or Amino Acid Sequence Disclosures
Summary of Requirements for Patent Applications Filed On Or After July 1, 2022, That Have Sequence Disclosures
37 CFR 1.831(a) requires that patent applications which contain disclosures of nucleotide and/or amino acid sequences that fall within the definitions of 37 CFR 1.831(b) must contain a “Sequence Listing XML”, 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 1.831-1.835. This “Sequence Listing XML” part of the disclosure may be submitted:
1. In accordance with 37 CFR 1.831(a) using the symbols and format requirements of 37 CFR 1.832 through 1.834 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”) in XML format, together with an incorporation by reference statement of the material in the XML file in a separate paragraph of the specification (an incorporation by reference paragraph) as required by 37 CFR 1.835(a)(2) or 1.835(b)(2) identifying:
a. the name of the XML file
b. the date of creation; and
c. the size of the XML file in bytes; or
2. In accordance with 37 CFR 1.831(a) using the symbols and format requirements of 37 CFR 1.832 through 1.834 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 statement of the material in the XML format according to 37 CFR 1.52(e)(8) and 37 CFR 1.835(a)(2) or 1.835(b)(2) in a separate paragraph of the specification identifying:
a. the name of the XML file;
b. the date of creation; and
c. the size of the XML file in bytes.
SPECIFIC DEFICIENCIES AND THE REQUIRED RESPONSE TO THIS NOTICE ARE AS FOLLOWS:
Specific deficiency - Sequences appearing in the specification are not identified by sequence identifiers (i.e., “SEQ ID NO:X” or the like) in accordance with 37 CFR 1.831(c), see the amino acid sequence disclosed at page 2
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.
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.
Claim 3 and claim 6 depending therefrom 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 applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claim 3 recites the limitation “d) one or more nucleotide sequence(s) which is/are comprised in the nucleotide sequence a), b) or C) and wherein said nucleotide sequence d) is fused in frame with the nucleotide sequences of a), b) and/or c)”. The metes and bounds of the limitation in the context of the above claims are not clear. It is unclear how the nucleotide sequence of d) is comprised in the nucleotide sequence of a), b), and/or c) and also fused in frame with the nucleotide sequence of a), b) and/or c). Clarification is requested.
Claim 5 is 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 applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
The term “gradually” in claim 5 is a relative term which renders the claim indefinite. The term “minimal” 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. It is unclear as to what degree of reduction in the level of the fusion protein is considered as “gradual”. A perusal of the specification did not provide a clear definition for the above limitation. Without a clear definition in terms of numerical value, those skilled in the art would be unable to ascertain what degree of reduction in the level of the fusion protein is gradual.
Claims 7, 9-10 and 13 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 applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claims 7, 9-10 and 13 recite the phrase “preferably”. The phrase renders the claims indefinite because it is unclear whether the limitation(s) following the phrase are part of the claimed invention. See MPEP § 2173.05(d).
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 and 5-20 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.
MPEP 2111.01 states that ''[d]uring examination, the claims must be interpreted as broadly as their terms reasonably allow.'' In the instant case, the limitations “an amino acid sequence” recited in claim 1 and “fragment” have been broadly interpreted as any amino acid sequence comprising as little as two contiguous amino acids of SEQ ID NO:1. Therefore, the claims are directed to an expression cassette, vector, or host cell comprising a polynucleotide encoding a fusion protein comprising (a) any polypeptide comprising as little as two contiguous amino acids of SEQ ID NO:1 or any polypeptide having at least 60% sequence identity to SEQ ID NO:1, wherein the polypeptide has the function directing protein decay and (b) a protein of interest. Thus, the claims are directed to an expression cassette, vector or host cell comprising a fusion protein comprising (a) a genus of polypeptides having unknown structure, wherein the polypeptide has the function directing protein decay, and (b) a protein of interest.
MPEP 2163 I. states that 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.
MPEP 2163. II.A.3.(a) sates that “Possession may be shown in many ways. For example, possession may be shown by describing an actual reduction to practice of the claimed invention. Possession may also be shown by a clear depiction of the invention in detailed drawings or in structural chemical formulas which permit a person skilled in the art to clearly recognize that inventor had possession of the claimed invention. An adequate written description of the invention may be shown by any description of sufficient, relevant, identifying characteristics so long as a person skilled in the art would recognize that the inventor had possession of the claimed invention.
According to MPEP 2163.II.A.3.(a).ii), “Satisfactory disclosure of a ‘representative number’ depends on whether one of skill in the art would recognize that the applicant was in possession of the necessary common attributes or features possessed by the members of the genus in view of the species disclosed. For inventions in an unpredictable art, adequate written description of a genus which embraces widely variant species cannot be achieved by disclosing only one species within the genus…Instead, the disclosure must adequately reflect the structural diversity of the claimed genus, either through the disclosure of sufficient species that are ‘representative of the full variety or scope of the genus,’ or by the establishment of ‘a reasonable structure-function correlation.’"
The recitation of “directs protein decay” fails to provide a sufficient description of the genus of the polypeptides as it merely describes the functional features of the genus without providing any definition of the structural features of the species within the genus. The specification does not specifically define any of the species that fall within the genus. The specification does not define any structural features commonly possessed by members of the genus that distinguish them from others. One skilled in the art therefore cannot, as one can do with a fully described genus, visualize or recognize the identity of the members of the genus.
The polypeptide comprising the amino acid sequence of SEQ ID NO:1 was not known in the prior art. A search of SEQ ID NO:1 did not result in polypeptides having any significant similarity, as shown below.
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Fransceus (J Ind Microbiol Biotechnol. 2017 May;44(4-5):687-695. – form PTO-892) reviews protein engineering techniques, such as random mutagenesis and recombination, directed evolution and iterative or combinatory saturation “hotspots”. Fransceus states that “a recurring problem, however, is choosing which amino acid positions should be targeted. Answering this question is not an easy feat and requires substantial insight in the relationship between an enzyme’s sequence or structure and its properties.” Sanavia (Computational and Structural Biotechnology Journal, Volume 18, 2020, Pages 1968-1979. – form PTO-892) discloses challenges in the prediction of protein stability in the occurrence of multiple mutations. “Multiple-point mutations are common variations of the protein sequence that may be needed in protein engineering when a single-point mutation is not enough to yield the desired stability change. Dealing with multiple-site variations adds another level of complexity beyond the prediction of the effect of a single variant on protein stability, since it requires the learning of many types of combinatorial effects”.
The specification is limited to description of an expression cassette, vector, or host cell comprising a polynucleotide encoding a fusion protein comprising (a) the polypeptide comprising the amino acid sequence of SEQ ID NO:1 and (b) a protein of interest, wherein the polypeptide of (a) directs decay of the protein of interest of (b). While MPEP 2163 acknowledges that in certain situations “one species adequately supports a genus,” it also acknowledges that “[f]or inventions in an unpredictable art, adequate written description of a genus which embraces widely variant species cannot be achieved by disclosing only one species within the genus.” In view of the widely variant species encompassed by the genus, the example described above is not enough and does not constitute a representative number of species to describe the whole genus. Therefore, the specification fails to describe a representative species of the claimed genus.
Further, one of skill in the art could identify fragments and polypeptides having at least 60% sequence identity to SEQ ID NO:1. However, there is no teaching regarding which amino acid or which 40% of the amino acids can vary from SEQ ID NO:1 and result in polypeptide having the function of directing protein decay. An important consideration is that structure is not necessarily a reliable indicator of function. In the instant case, there is no disclosure relating similarity of structure to conservation of function. Conservation of structure is not necessarily a surrogate for conservation of function.
Given this lack of description of the representative species encompassed by the genus of the claims, the specification fails to sufficiently describe the claimed invention in such full, clear, concise, and exact terms that a skilled artisan would recognize that applicants were in possession of the inventions of claims 1-3 and 5-20
Claims 1-3 and 5-20 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, because the specification, while being enabling for an expression cassette, vector, or host cell comprising a polynucleotide encoding a fusion protein comprising (a) the polypeptide comprising the amino acid sequence of SEQ ID NO:1 and (b) a protein of interest, wherein the polypeptide of (a) directs decay of the protein of interest of (b), does not reasonably provide enablement an expression cassette, vector, or host cell comprising a polynucleotide encoding a fusion protein comprising (a) any polypeptide having unknown structure and having the function of directing protein decay and (b) a protein of interest. The specification does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the invention commensurate in scope with these claims.
Factors to be considered in determining whether undue experimentation is required are summarized in In re Wands 858 F.2d 731, 8 USPQ2nd 1400 (Fed. Cir, 1988). They include (1) the quantity of experimentation necessary, (2) the amount of direction or guidance presented, (3) the presence or absence of working examples, (4) the nature of the invention, (5) the state of the prior art, (6) the relative skill of those in the art, (7) the predictability or unpredictability of the art, and (8) the breadth of the claims.
The breadth of the claims.
MPEP 2111.01 states that ''[d]uring examination, the claims must be interpreted as broadly as their terms reasonably allow.'' In the instant case, the limitations “an amino acid sequence” recited in claim 1 and “fragment” have been broadly interpreted as any amino acid sequence comprising as little as two contiguous amino acids of SEQ ID NO:1. Therefore, the claims are directed to an expression cassette, vector, or host cell comprising a polynucleotide encoding a fusion protein comprising (a) any polypeptide comprising as little as two contiguous amino acids of SEQ ID NO:1 or any polypeptide having at least 60% sequence identity to SEQ ID NO:1, wherein the polypeptide has the function of directing protein decay and (b) a protein of interest. Thus, the claims are directed to an expression cassette, vector or host cell comprising a fusion protein comprising (a) a polypeptide having unknown structure, wherein the polypeptide has the function of directing protein decay and (b) a protein of interest.
The claims are not commensurate with the enablement provided by the disclosure with regard to the extremely large number of polypeptides having the function of directing protein decay. In the instant case, the specification is limited to an expression cassette, vector, or host cell comprising a polynucleotide encoding a fusion protein comprising (a) the polypeptide comprising the amino acid sequence of SEQ ID NO:1 and (b) a protein of interest, wherein the polypeptide of (a) directs decay of the protein of interest of (b).
The quantity of experimentation required to practice the claimed invention based on the teachings of the specification.
While protein isolation techniques, recombinant and mutagenesis techniques were known in the art at the time of the invention, e.g. mutagenesis, and it is routine in the art to screen for variants comprising multiple substitutions or multiple modifications as encompassed by the instant claims, the specific amino acid positions within the protein's sequence where amino acid modifications can be made with a reasonable expectation of success in obtaining the desired activity/utility are limited in any protein and the result of such modifications is unpredictable. In addition, one skilled in the art would expect any tolerance to modification for a given protein to diminish with each further and additional modification, e.g. multiple substitutions.
In the absence of: (a) rational and predictable scheme for making any polypeptide comprising as little as two contiguous amino acids of SEQ ID NO:1 or having at least 60% sequence identity to SEQ ID NO:1, wherein the polypeptide has the function of directing protein decay and (b) a correlation between structure and the function of directing protein decay, the specification provides insufficient guidance as to which of the essentially infinite possible choices is likely to be successful. One of skill in the art would have to test these infinite possible polypeptides to determine which polypeptides have the function of directing protein decay. While enablement is not precluded by the necessity for routine screening, if a large amount of screening is required, as is the case herein, the specification must provide a reasonable amount of guidance which respect to the direction in which the experimentation should proceed so that a reasonable number of species can be selected for testing. In view of the fact that such guidance has not been provided in the instant specification, it would require undue experimentation to enable the full scope of the claims.
The state of prior art, the relative skill of those in the art, and predictability or unpredictability of the art.
Since the amino acid sequence of the protein determines its structural and functional properties, predictability of which changes can be tolerated in a protein's amino acid sequence and obtain the desired activity requires a knowledge of and guidance with regard to which amino acids in the protein's sequence, if any, are tolerant of modification and which are conserved (i.e. expectedly intolerant to modification), and detailed knowledge of the ways in which the proteins' structure relates to its function. In the instant case, neither the specification or the art provide a correlation between structure and activity such that one of skill in the art can envision the structure of any polypeptides having the function of directing protein decay or predict said function of a polypeptide from its primary structure. In addition, the art does not provide any teaching or guidance as to (1) which amino acids within the polynucleotide of SEQ ID NO:1 or any fragment thereof that can be modified and which ones are conserved such that one of skill in the art can make the recited polypeptides having the function of directing protein decay, (2) which segments of the polypeptide of SEQ ID NO:1 that are essential for polypeptides having the function of directing protein decay, and (3) the general tolerance of the polypeptide of SEQ ID NO:1 to structural modifications and the extent of such tolerance. The art clearly teaches that changes in a protein's amino acid sequence to obtain the desired activity without any guidance/knowledge as to which amino acids in a protein are required for that activity is highly unpredictable. At the time of the invention there was a high level of unpredictability associated with altering a polypeptide sequence with an expectation that the polypeptide will maintain the desired activity. For example, Studer (Residue mutations and their impact on protein structure and function: detecting beneficial and pathogenic changes. Biochem. J. (2013) 449, 581–594. – form PTO-892) teach that (1) protein engineers are frequently surprised by the range of effects caused by single mutations that they hoped would change only one specific and simple property in enzymes, (2) the often surprising results obtained by experiments where single mutations are made reveal how little is known about the rules of protein stability, and (3) the difficulties in designing de novo stable proteins with specific functions.
The polypeptide comprising the amino acid sequence of SEQ ID NO:1 was not known in the prior art. A search of SEQ ID NO:1 did not result in polypeptides having any significant similarity, as shown below.
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Fransceus (J Ind Microbiol Biotechnol. 2017 May;44(4-5):687-695. – form PTO-892) reviews protein engineering techniques, such as random mutagenesis and recombination, directed evolution and iterative or combinatory saturation “hotspots”. Fransceus states that “a recurring problem, however, is choosing which amino acid positions should be targeted. Answering this question is not an easy feat and requires substantial insight in the relationship between an enzyme’s sequence or structure and its properties.” Sanavia (Computational and Structural Biotechnology Journal, Volume 18, 2020, Pages 1968-1979. – form PTO-892) discloses challenges in the prediction of protein stability in the occurrence of multiple mutations. “Multiple-point mutations are common variations of the protein sequence that may be needed in protein engineering when a single-point mutation is not enough to yield the desired stability change. Dealing with multiple-site variations adds another level of complexity beyond the prediction of the effect of a single variant on protein stability, since it requires the learning of many types of combinatorial effects”.
The amount of direction or guidance presented and the existence of working examples.
The specification is limited to an expression cassette, vector, or host cell comprising a polynucleotide encoding a fusion protein comprising (a) the polypeptide comprising the amino acid sequence of SEQ ID NO:1 and (b) a protein of interest, wherein the polypeptide of (a) directs decay of the protein of interest of (b). However, the speciation fails to provide any information as to (1) specific substrates associated with polypeptides having at least 60% sequence identity to SEQ ID NO: 1 or fragments thereof or (2) structural elements required in a polypeptide having the function of directing protein decay. No correlation between structure and function of having the function of directing protein decay has been presented. There is no information or guidance as to which amino acid residues in the polypeptide of SEQ ID NO: 1 that can be modified and which ones are to be conserved to create a polypeptide having the function of directing protein decay.
Thus, in view of the overly broad scope of the claims, the lack of guidance and working examples provided in the specification, the high level of unpredictability of the prior art in regard to structural changes and their effect on function and the lack of knowledge about a correlation between structure and function, an undue experimentation would be necessary one having ordinary skill in the art to make and use the claimed invention in a manner reasonably correlated with the scope of the claims. The scope of the claims must bear a reasonable correlation with the scope of enablement (In re Fisher, 166 USPQ 19 24 (CCPA 1970)). Without sufficient guidance, determination of polypeptides having the desired biological characteristics recited in the claims are unpredictable and the experimentation left to those skilled in the art is unnecessarily, and improperly, extensive and undue. See In re Wands 858 F.2d 731, 8 USPQ2nd 1400 (Fed. Cir, 1988).
Claim Rejections - 35 USC § 102
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.
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.
Claim(s) 1-3, 5-8, 11-12, 15-16, and 19 is/are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Lin (US 10,550,379 - form PTO-1449 or WO 2017/004022 – form PTO-892. US 10,550,379 is used for specific passages of Lin).
MPEP 2111.01 states that ''[d]uring examination, the claims must be interpreted as broadly as their terms reasonably allow.'' The limitations “an amino acid sequence” recited in claim 1 and “fragment” have been broadly interpreted as any amino acid sequence comprising as little as two contiguous amino acids of SEQ ID NO:1.
Regarding claim 1, Lin discloses an expression cassette (Column 18, line 64 through Column 19, line 12) encoding a fusion protein, comprising a) a nucleotide sequence encoding (i) a degron comprising the amino acid sequence shown in SEQ ID NO: 1, directs protein decay, and b) a nucleotide sequence encoding a protein of interest, wherein nucleotide sequence a) and b) are fused together in frame (Column 1, lines 49-64 and claims 1 and 21). The degron of SEQ ID NO:1 of Lin comprises at least two contiguous amino acids of SEQ ID NO:1 of the instant application (see the sequence alignment below). The degron of SEQ ID NO:1 of Lin is 42 amino acids long and therefore reads on a fragment of at least 18 amino acid long comprising as little as two contiguous amino acids of SEQ ID NO:1 of the instant application (see the sequence alignment below).
Regarding claim 2, the degron of Lin is located at the N-terminus or at the C-terminus (Column 2, lines 28-63 and claim 2).
Regarding claim 3, the expression cassette further comprises a nucleotide sequence encoding a cleavable linker fused to the 5'- and/ or 3'-end of the nucleotide sequence encoding the fusion protein (Column 2, lines 28-63 and claim 1).
Regarding claim 5, the level of the fusion protein reduces when expressed in a cell in comparison to a cell which expresses the protein of interest (abstract).
Regarding claim 6, the expression cassette of Lin comprises at least 3 nucleotides and encodes a heterologous polypeptide, wherein said heterologous polypeptide is a linker cleavable by a protease (abstract and claim 1).
Regarding claim 7, the expression cassette of Lin has uses various promoters such as a the constitutive CMV promoter (Column 30, lines 55-65).
Regarding claim 8, the protein of interest expression cassette of Lin encodes a polypeptide selected from a group consisting of enzymes, receptors, hormones, membrane proteins, and membrane associated proteins (claim 18).
Regarding claim 11, Lin discloses a vector comprising the expression cassette (Column 3, lines 29-39).
Regarding claim 12, Lin discloses a host cell comprising the expression cassette (claim 28).
Regarding claim 15, Lin discloses a mammalian host cell (Column 3, lines 53-58 and claim 29)
Regarding claim 16, the host cell of Lin not express one or more sterol acyltransferases of SEQ ID NO:15 and/or 16 of the instant application.
Regarding claim 19, the host cell of Lin comprises a vector comprising the expression cassette (Column 3, line 33 through Column 4, line 4).
Therefore, the reference of Lin anticipates claims 1-3, 5-8, 11-12, 15-16, and 19.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claim(s) 1-3, 5-9, and 11-20 is/are rejected under 35 U.S.C. 103 as being unpatentable over Lin (US 10,550,379 - form PTO-1449 or WO 2017/004022 – form PTO-892. US 10,550,379 is used for specific passages of Lin) and Schulze (WO 2019/197327 – form PTO-892).
MPEP 2111.01 states that ''[d]uring examination, the claims must be interpreted as broadly as their terms reasonably allow.'' The limitations “an amino acid sequence” recited in claim 1 and “fragment” have been broadly interpreted as any amino acid sequence comprising as little as two contiguous amino acids of SEQ ID NO:1.
Regarding claim 1, Lin discloses an expression cassette (Column 18, line 64 through Column 19, line 12) encoding a fusion protein, comprising a) a nucleotide sequence encoding (i) a degron comprising the amino acid sequence shown in SEQ ID NO: 1, directs protein decay, and b) a nucleotide sequence encoding a protein of interest, wherein nucleotide sequence a) and b) are fused together in frame (Column 1, lines 49-64 and claims 1 and 21). The degron of SEQ ID NO:1 of Lin comprises at least two contiguous amino acids of SEQ ID NO:1 of the instant application (see the sequence alignment below). The degron of SEQ ID NO:1 of Lin is 42 amino acids long and therefore reads on a fragment of at least 18 amino acid long comprising as little as two contiguous amino acids of SEQ ID NO:1 of the instant application (see the sequence alignment below).
Regarding claim 2, the degron of Lin is located at the N-terminus or at the C-terminus (Column 2, lines 28-63 and claim 2).
Regarding claim 3, the expression cassette further comprises a nucleotide sequence encoding a cleavable linker fused to the 5'- and/ or 3'-end of the nucleotide sequence encoding the fusion protein (Column 2, lines 28-63 and claim 1).
Regarding claim 5, the level of the fusion protein reduces when expressed in a cell in comparison to a cell which expresses the protein of interest (abstract).
Regarding claim 6, the expression cassette of Lin comprises at least 3 nucleotides and encodes a heterologous polypeptide, wherein said heterologous polypeptide is a linker cleavable by a protease (abstract and claim 1).
Regarding claim 7, the expression cassette of Lin has uses various promoters such as a the constitutive CMV promoter (Column 30, lines 55-65).
Regarding claim 8, the protein of interest expression cassette of Lin encodes a polypeptide selected from a group consisting of enzymes, receptors, hormones, membrane proteins, and membrane associated proteins (claim 18).
Regarding claim 11, Lin discloses a vector comprising the expression cassette (Column 3, lines 29-39).
Regarding claim 12, Lin discloses a host cell comprising the expression cassette (claim 28).
Regarding claim 15, Lin discloses a mammalian host cell (Column 3, lines 53-58 and claim 29)
Regarding claim 16, the host cell of Lin not express one or more sterol acyltransferases of SEQ ID NO:15 and/or 16 of the instant application.
Regarding claim 19, the host cell of Lin comprises a vector comprising the expression cassette (Column 3, line 33 through Column 4, line 4).
However, Lin does not disclose a fusion protein wherein the protein of interest is a lanosterol synthase (ERG7) having the amino acid sequence of SEQ ID NO:3 and encoded by SEQ ID NO:4 nor a method of producing a triterpenoid.
Regarding claims 9 and 13-14, Schulze discloses an engineered host cell to repress the lanosterol synthase (ERG7) having the amino acid sequence of SEQ ID NO:9 and a polynucleotide encoding said lanosterol synthase ([0015] and claim 4). The lanosterol synthase of SEQ ID NO:9 of Schulze is identical to the lanosterol synthase of SEQ ID NO:3 of the instant application and the polynucleotide encoding the lanosterol synthase of SEQ ID NO:9 of Schulze is identical to the polynucleotide of SEQ ID NO:4 of the instant application (see the sequence alignments below).
Regarding claims 15 and 20, Schulze discloses a yeast host engineered to repress the lanosterol synthase (claims 1 and 10).
Regarding claim 16, the yeast host cell of Schulze does not express one or more sterol acyltransferases of SEQ ID NO:15 and/or 16 of the instant application.
Regarding claim 17, Schulze discloses using a cytochrome P450 reductase and a cytochrome P450 monooxygenase to produce triterpenes in yeast ([006]).
Regarding claim 18, Schulze discloses a method of increasing the yield of a triterpenoid in a host cell or yeast host cell, wherein the lanosterol synthase (ERG7) is repressed (claims 1 and 4).
Therefore, in combining the teachings of Lin and Schulze, it would have been obvious to one having ordinary skill in the art before the time the claimed invention was effectively filed to make an expression cassette, vector, or yeast host cell comprising a polynucleotide encoding a fusion protein comprising (a) the degron of Lin and (b) lanosterol synthase (ERG7), and a cleavable linker, and a method of producing a triterpenoid using said yeast host cell. One of ordinary skill in the art at the time the invention was made would have been motivated to do so in order to control the enzymatic activity of lanosterol synthase to increase the yield of triterpenoid production. One of ordinary skill in the art would have had a reasonable expectation of success since Lin discloses a polynucleotide encoding a fusion protein comprising a degron and a protein of interest, wherein the degron decays the protein of interest and Schulze discloses repressing lanosterol synthase (ERG7) in order to increase the yield of triterpenoid production.
Therefore, the above references render claims 1-3, 5-9, and 11-20 prima facie obvious.
Conclusion
Claims1-20 are pending.
Claims 1-3 and 5-20 are rejected.
Claim 4 is objected to as being dependent upon a rejected base claim, but would be allowable if rewritten in independent form including all of the limitations of the base claim and any intervening claims.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to YONG D PAK whose telephone number is (571)272-0935. The examiner can normally be reached M-Th: 5:30 am - 3:30 pm.
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If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Robert Mondesi can be reached on 408-918-7584. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
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/YONG D PAK/Primary Examiner, Art Unit 1652
Sequence alignment between the peptide of SEQ ID NO:1 of the instant application (“Qy”) and the degron of SEQ ID NO:1 of Lin (“Db”)
Title: US-18-709-148A-1
Perfect score: 155
Sequence: 1 LRLEQVLVLVLEQFCLKVKNYSLVLSQFWLN 31
Scoring table: BLOSUM62
Gapop 10.0 , Gapext 0.5
Searched: 1 seqs, 42 residues
Total number of hits satisfying chosen parameters: 1
Minimum DB seq length: 0
Maximum DB seq length: inf
Post-processing: Minimum Match 0%
Maximum Match 100%
Listing first 1 summaries
Database : AASEQ2_03172026_154334.pep:*
SUMMARIES
%
Result Query
No. Score Match Length DB ID Description
----------------------------------------------------------------------------
1 19 12.3 42 1 AASEQ2_03172026_154334
ALIGNMENTS
RESULT 1
AASEQ2_03172026_154334
Query Match 12.3%; Score 19; DB 1; Length 42;
Best Local Similarity 30.0%;
Matches 3; Conservative 3; Mismatches 4; Indels 0; Gaps 0;
Qy 7 LVLVLEQFCL 16
:: | :||
Db 31 VLAALAAYCL 40
Sequence alignment between the lanosterol synthase (ERG7) of SEQ ID NO:3 of the instant application (“Qy”) and the lanosterol synthase (ERG7) of SEQ ID NO:9 of Schulze (“Db”)
BGW52013
ID BGW52013 standard; DNA; 2196 BP.
XX
AC BGW52013;
XX
DT 12-DEC-2019 (first entry)
XX
DE Saccharomyces cerevisiae lanosterol synthase (ERG7) DNA, SEQ ID 20.
XX
KW ERG7; ds; expression; gene; genetic engineering; lanosterol synthase.
XX
OS Saccharomyces cerevisiae.
XX
FH Key Location/Qualifiers
FT CDS 1..2196
FT /*tag= a
FT /product= " lanosterol synthase (ERG7) protein"
XX
CC PN WO2019197327-A1.
XX
CC PD 17-OCT-2019.
XX
CC PF 08-APR-2019; 2019WO-EP058789.
XX
PR 09-APR-2018; 2018EP-00166374.
XX
CC PA (FRAU ) FRAUNHOFER GES FOERDERUNG ANGEWANDTEN EV.
CC PA (UYMU ) UNIV MUENSTER WESTFAELISCHE WILHELMS.
XX
CC PI Schulze Gronover C, Boje Mueller LG, Van Deenen N, Broeker JN;
XX
DR WPI; 2019-86338C/83.
DR P-PSDB; BGW52002.
XX
CC PT Increasing yield of one of oxidosqualene, triterpenes and/or
CC PT triterpenoids in host cell involves engineering host cell to overexpress
CC PT a 3-hydroxy-3-methylglutaryl-coenzyme A reductase comprising an amino
CC PT acids.
XX
CC PS Example 2; SEQ ID NO 20; 96pp; English.
XX
CC The invention relates to a novel method of increasing the yield of at
CC least one of oxidosqualene, triterpenes and triterpenoids in a host cell.
CC The method involves engineering host cell to overexpress a 3-hydroxy-3-
CC methylglutaryl-coenzyme A reductase comprising an amino acid shown in SEQ
CC ID 1 (see BGW51994). The invention also provides a recombinant host cell
CC for manufacturing one of oxidosqualene, triterpenes and triterpenoids.
CC The method is useful for manufacturing one of oxidosqualene, triterpenes
CC and triterpenoids.
XX
SQ Sequence 2196 BP; 652 A; 448 C; 476 G; 620 T; 0 U; 0 Other;
Query Match 100.0%; Score 3939; Length 731;
Best Local Similarity 100.0%;
Matches 731; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MTEFYSDTIGLPKTDPRLWRLRTDELGRESWEYLTPQQAANDPPSTFTQWLLQDPKFPQP 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MTEFYSDTIGLPKTDPRLWRLRTDELGRESWEYLTPQQAANDPPSTFTQWLLQDPKFPQP 60
Qy 61 HPERNKHSPDFSAFDACHNGASFFKLLQEPDSGIFPCQYKGPMFMTIGYVAVNYIAGIEI 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 HPERNKHSPDFSAFDACHNGASFFKLLQEPDSGIFPCQYKGPMFMTIGYVAVNYIAGIEI 120
Qy 121 PEHERIELIRYIVNTAHPVDGGWGLHSVDKSTVFGTVLNYVILRLLGLPKDHPVCAKARS 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 PEHERIELIRYIVNTAHPVDGGWGLHSVDKSTVFGTVLNYVILRLLGLPKDHPVCAKARS 180
Qy 181 TLLRLGGAIGSPHWGKIWLSALNLYKWEGVNPAPPETWLLPYSLPMHPGRWWVHTRGVYI 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 TLLRLGGAIGSPHWGKIWLSALNLYKWEGVNPAPPETWLLPYSLPMHPGRWWVHTRGVYI 240
Qy 241 PVSYLSLVKFSCPMTPLLEELRNEIYTKPFDKINFSKNRNTVCGVDLYYPHSTTLNIANS 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 PVSYLSLVKFSCPMTPLLEELRNEIYTKPFDKINFSKNRNTVCGVDLYYPHSTTLNIANS 300
Qy 301 LVVFYEKYLRNRFIYSLSKKKVYDLIKTELQNTDSLCIAPVNQAFCALVTLIEEGVDSEA 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 LVVFYEKYLRNRFIYSLSKKKVYDLIKTELQNTDSLCIAPVNQAFCALVTLIEEGVDSEA 360
Qy 361 FQRLQYRFKDALFHGPQGMTIMGTNGVQTWDCAFAIQYFFVAGLAERPEFYNTIVSAYKF 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 FQRLQYRFKDALFHGPQGMTIMGTNGVQTWDCAFAIQYFFVAGLAERPEFYNTIVSAYKF 420
Qy 421 LCHAQFDTECVPGSYRDKRKGAWGFSTKTQGYTVADCTAEAIKAIIMVKNSPVFSEVHHM 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 LCHAQFDTECVPGSYRDKRKGAWGFSTKTQGYTVADCTAEAIKAIIMVKNSPVFSEVHHM 480
Qy 481 ISSERLFEGIDVLLNLQNIGSFEYGSFATYEKIKAPLAMETLNPAEVFGNIMVEYPYVEC 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 ISSERLFEGIDVLLNLQNIGSFEYGSFATYEKIKAPLAMETLNPAEVFGNIMVEYPYVEC 540
Qy 541 TDSSVLGLTYFHKYFDYRKEEIRTRIRIAIEFIKKSQLPDGSWYGSWGICFTYAGMFALE 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 TDSSVLGLTYFHKYFDYRKEEIRTRIRIAIEFIKKSQLPDGSWYGSWGICFTYAGMFALE 600
Qy 601 ALHTVGETYENSSTVRKGCDFLVSKQMKDGGWGESMKSSELHSYVDSEKSLVVQTAWALI 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 601 ALHTVGETYENSSTVRKGCDFLVSKQMKDGGWGESMKSSELHSYVDSEKSLVVQTAWALI 660
Qy 661 ALLFAEYPNKEVIDRGIDLLKNRQEESGEWKFESVEGVFNHSCAIEYPSYRFLFPIKALG 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 661 ALLFAEYPNKEVIDRGIDLLKNRQEESGEWKFESVEGVFNHSCAIEYPSYRFLFPIKALG 720
Qy 721 MYSRAYETHTL 731
|||||||||||
Db 721 MYSRAYETHTL 731
Sequence alignment between the polynucleotide SEQ ID NO:4 of the instant application (“Qy”) and the polynucleotide encoding the lanosterol synthase (ERG7) of SEQ ID NO:9 of Schulze (“Db”)
BGW52013
ID BGW52013 standard; DNA; 2196 BP.
XX
AC BGW52013;
XX
DT 12-DEC-2019 (first entry)
XX
DE Saccharomyces cerevisiae lanosterol synthase (ERG7) DNA, SEQ ID 20.
XX
KW ERG7; ds; expression; gene; genetic engineering; lanosterol synthase.
XX
OS Saccharomyces cerevisiae.
XX
FH Key Location/Qualifiers
FT CDS 1..2196
FT /*tag= a
FT /product= " lanosterol synthase (ERG7) protein"
XX
CC PN WO2019197327-A1.
XX
CC PD 17-OCT-2019.
XX
CC PF 08-APR-2019; 2019WO-EP058789.
XX
PR 09-APR-2018; 2018EP-00166374.
XX
CC PA (FRAU ) FRAUNHOFER GES FOERDERUNG ANGEWANDTEN EV.
CC PA (UYMU ) UNIV MUENSTER WESTFAELISCHE WILHELMS.
XX
CC PI Schulze Gronover C, Boje Mueller LG, Van Deenen N, Broeker JN;
XX
DR WPI; 2019-86338C/83.
DR P-PSDB; BGW52002.
XX
CC PT Increasing yield of one of oxidosqualene, triterpenes and/or
CC PT triterpenoids in host cell involves engineering host cell to overexpress
CC PT a 3-hydroxy-3-methylglutaryl-coenzyme A reductase comprising an amino
CC PT acids.
XX
CC PS Example 2; SEQ ID NO 20; 96pp; English.
XX
CC The invention relates to a novel method of increasing the yield of at
CC least one of oxidosqualene, triterpenes and triterpenoids in a host cell.
CC The method involves engineering host cell to overexpress a 3-hydroxy-3-
CC methylglutaryl-coenzyme A reductase comprising an amino acid shown in SEQ
CC ID 1 (see BGW51994). The invention also provides a recombinant host cell
CC for manufacturing one of oxidosqualene, triterpenes and triterpenoids.
CC The method is useful for manufacturing one of oxidosqualene, triterpenes
CC and triterpenoids.
XX
SQ Sequence 2196 BP; 652 A; 448 C; 476 G; 620 T; 0 U; 0 Other;
Query Match 100.0%; Score 2196; Length 2196;
Best Local Similarity 100.0%;
Matches 2196; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 ATGACAGAATTTTATTCTGACACAATCGGTCTACCAAAGACAGATCCACGTCTTTGGAGA 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 ATGACAGAATTTTATTCTGACACAATCGGTCTACCAAAGACAGATCCACGTCTTTGGAGA 60
Qy 61 CTGAGAACTGATGAGCTAGGCCGAGAAAGCTGGGAATATTTAACCCCTCAGCAAGCCGCA 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 CTGAGAACTGATGAGCTAGGCCGAGAAAGCTGGGAATATTTAACCCCTCAGCAAGCCGCA 120
Qy 121 AACGACCCACCATCCACTTTCACGCAGTGGCTTCTTCAAGATCCCAAATTTCCTCAACCT 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 AACGACCCACCATCCACTTTCACGCAGTGGCTTCTTCAAGATCCCAAATTTCCTCAACCT 180
Qy 181 CATCCAGAAAGAAATAAGCATTCACCAGATTTTTCAGCCTTCGATGCGTGTCATAATGGT 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 CATCCAGAAAGAAATAAGCATTCACCAGATTTTTCAGCCTTCGATGCGTGTCATAATGGT 240
Qy 241 GCATCTTTTTTCAAACTGCTTCAAGAGCCTGACTCAGGTATTTTTCCGTGTCAATATAAA 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 GCATCTTTTTTCAAACTGCTTCAAGAGCCTGACTCAGGTATTTTTCCGTGTCAATATAAA 300
Qy 301 GGACCCATGTTCATGACAATCGGTTACGTAGCCGTAAACTATATCGCCGGTATTGAAATT 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 GGACCCATGTTCATGACAATCGGTTACGTAGCCGTAAACTATATCGCCGGTATTGAAATT 360
Qy 361 CCTGAGCATGAGAGAATAGAATTAATTAGATACATCGTCAATACAGCACATCCGGTTGAT 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 CCTGAGCATGAGAGAATAGAATTAATTAGATACATCGTCAATACAGCACATCCGGTTGAT 420
Qy 421 GGTGGCTGGGGTCTACATTCTGTTGACAAATCCACCGTGTTTGGTACAGTATTGAACTAT 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 GGTGGCTGGGGTCTACATTCTGTTGACAAATCCACCGTGTTTGGTACAGTATTGAACTAT 480
Qy 481 GTAATCTTACGTTTATTGGGTCTACCCAAGGACCACCCGGTTTGCGCCAAGGCAAGAAGC 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 GTAATCTTACGTTTATTGGGTCTACCCAAGGACCACCCGGTTTGCGCCAAGGCAAGAAGC 540
Qy 541 ACATTGTTAAGGTTAGGCGGTGCTATTGGATCCCCTCACTGGGGAAAAATTTGGCTAAGT 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 ACATTGTTAAGGTTAGGCGGTGCTATTGGATCCCCTCACTGGGGAAAAATTTGGCTAAGT 600
Qy 601 GCACTAAACTTGTATAAATGGGAAGGTGTGAACCCTGCCCCTCCTGAAACTTGGTTACTT 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 601 GCACTAAACTTGTATAAATGGGAAGGTGTGAACCCTGCCCCTCCTGAAACTTGGTTACTT 660
Qy 661 CCATATTCACTGCCCATGCATCCGGGGAGATGGTGGGTTCATACTAGAGGTGTTTACATT 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 661 CCATATTCACTGCCCATGCATCCGGGGAGATGGTGGGTTCATACTAGAGGTGTTTACATT 720
Qy 721 CCGGTCAGTTACCTGTCATTGGTCAAATTTTCTTGCCCAATGACTCCTCTTCTTGAAGAA 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 721 CCGGTCAGTTACCTGTCATTGGTCAAATTTTCTTGCCCAATGACTCCTCTTCTTGAAGAA 780
Qy 781 CTGAGGAATGAAATTTACACTAAACCGTTTGACAAGATTAACTTCTCCAAGAACAGGAAT 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 781 CTGAGGAATGAAATTTACACTAAACCGTTTGACAAGATTAACTTCTCCAAGAACAGGAAT 840
Qy 841 ACCGTATGTGGAGTAGACCTATATTACCCCCATTCTACTACTTTGAATATTGCGAACAGC 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 841 ACCGTATGTGGAGTAGACCTATATTACCCCCATTCTACTACTTTGAATATTGCGAACAGC 900
Qy 901 CTTGTAGTATTTTACGAAAAATACCTAAGAAACCGGTTCATTTACTCTCTATCCAAGAAG 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 901 CTTGTAGTATTTTACGAAAAATACCTAAGAAACCGGTTCATTTACTCTCTATCCAAGAAG 960
Qy 961 AAGGTTTATGATCTAATCAAAACGGAGTTACAGAATACTGATTCCTTGTGTATAGCACCT 1020
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 961 AAGGTTTATGATCTAATCAAAACGGAGTTACAGAATACTGATTCCTTGTGTATAGCACCT 1020
Qy 1021 GTTAACCAGGCGTTTTGCGCACTTGTCACTCTTATTGAAGAAGGGGTAGACTCGGAAGCG 1080
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1021 GTTAACCAGGCGTTTTGCGCACTTGTCACTCTTATTGAAGAAGGGGTAGACTCGGAAGCG 1080
Qy 1081 TTCCAGCGTCTCCAATATAGGTTCAAGGATGCATTGTTCCATGGTCCACAGGGTATGACC 1140
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1081 TTCCAGCGTCTCCAATATAGGTTCAAGGATGCATTGTTCCATGGTCCACAGGGTATGACC 1140
Qy 1141 ATTATGGGAACAAATGGTGTGCAAACCTGGGATTGTGCGTTTGCCATTCAATACTTTTTC 1200
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1141 ATTATGGGAACAAATGGTGTGCAAACCTGGGATTGTGCGTTTGCCATTCAATACTTTTTC 1200
Qy 1201 GTCGCAGGCCTCGCAGAAAGACCTGAATTCTATAACACAATTGTCTCTGCCTATAAATTC 1260
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1201 GTCGCAGGCCTCGCAGAAAGACCTGAATTCTATAACACAATTGTCTCTGCCTATAAATTC 1260
Qy 1261 TTGTGTCATGCTCAATTTGACACCGAGTGCGTTCCAGGTAGTTATAGGGATAAGAGAAAG 1320
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1261 TTGTGTCATGCTCAATTTGACACCGAGTGCGTTCCAGGTAGTTATAGGGATAAGAGAAAG 1320
Qy 1321 GGGGCTTGGGGCTTCTCAACAAAAACACAGGGCTATACAGTGGCAGATTGCACTGCAGAA 1380
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1321 GGGGCTTGGGGCTTCTCAACAAAAACACAGGGCTATACAGTGGCAGATTGCACTGCAGAA 1380
Qy 1381 GCAATTAAAGCCATCATCATGGTGAAAAACTCTCCCGTCTTTAGTGAAGTACACCATATG 1440
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1381 GCAATTAAAGCCATCATCATGGTGAAAAACTCTCCCGTCTTTAGTGAAGTACACCATATG 1440
Qy 1441 ATTAGCAGTGAACGTTTATTTGAAGGCATTGATGTGTTATTGAACCTACAAAACATCGGA 1500
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1441 ATTAGCAGTGAACGTTTATTTGAAGGCATTGATGTGTTATTGAACCTACAAAACATCGGA 1500
Qy 1501 TCTTTTGAATATGGTTCCTTTGCAACCTATGAAAAAATCAAGGCCCCACTAGCAATGGAA 1560
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1501 TCTTTTGAATATGGTTCCTTTGCAACCTATGAAAAAATCAAGGCCCCACTAGCAATGGAA 1560
Qy 1561 ACCTTGAATCCTGCTGAAGTTTTTGGTAACATAATGGTAGAATACCCATACGTGGAATGT 1620
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1561 ACCTTGAATCCTGCTGAAGTTTTTGGTAACATAATGGTAGAATACCCATACGTGGAATGT 1620
Qy 1621 ACTGATTCATCCGTTCTGGGGTTGACATATTTTCACAAGTACTTCGACTATAGGAAAGAG 1680
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1621 ACTGATTCATCCGTTCTGGGGTTGACATATTTTCACAAGTACTTCGACTATAGGAAAGAG 1680
Qy 1681 GAAATACGTACACGCATCAGAATCGCCATCGAATTCATAAAAAAATCTCAATTACCAGAT 1740
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1681 GAAATACGTACACGCATCAGAATCGCCATCGAATTCATAAAAAAATCTCAATTACCAGAT 1740
Qy 1741 GGAAGTTGGTATGGAAGCTGGGGTATTTGTTTTACATATGCCGGTATGTTTGCATTGGAG 1800
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1741 GGAAGTTGGTATGGAAGCTGGGGTATTTGTTTTACATATGCCGGTATGTTTGCATTGGAG 1800
Qy 1801 GCATTACACACCGTGGGGGAGACCTATGAGAATTCCTCAACGGTAAGAAAAGGTTGCGAC 1860
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1801 GCATTACACACCGTGGGGGAGACCTATGAGAATTCCTCAACGGTAAGAAAAGGTTGCGAC 1860
Qy 1861 TTCTTGGTCAGTAAACAGATGAAGGATGGCGGTTGGGGGGAATCAATGAAGTCCAGTGAA 1920
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1861 TTCTTGGTCAGTAAACAGATGAAGGATGGCGGTTGGGGGGAATCAATGAAGTCCAGTGAA 1920
Qy 1921 TTACATAGTTATGTGGATAGTGAAAAATCGCTAGTCGTTCAAACCGCATGGGCGCTAATT 1980
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1921 TTACATAGTTATGTGGATAGTGAAAAATCGCTAGTCGTTCAAACCGCATGGGCGCTAATT 1980
Qy 1981 GCACTTCTTTTCGCTGAATATCCTAATAAAGAAGTCATCGACCGCGGTATTGACCTTTTA 2040
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1981 GCACTTCTTTTCGCTGAATATCCTAATAAAGAAGTCATCGACCGCGGTATTGACCTTTTA 2040
Qy 2041 AAAAATAGACAAGAAGAATCCGGGGAATGGAAATTTGAAAGTGTAGAAGGTGTTTTCAAC 2100
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2041 AAAAATAGACAAGAAGAATCCGGGGAATGGAAATTTGAAAGTGTAGAAGGTGTTTTCAAC 2100
Qy 2101 CACTCTTGTGCAATTGAATACCCAAGTTATCGATTCTTATTCCCTATTAAGGCATTAGGT 2160
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2101 CACTCTTGTGCAATTGAATACCCAAGTTATCGATTCTTATTCCCTATTAAGGCATTAGGT 2160
Qy 2161 ATGTACAGCAGGGCATATGAAACACATACGCTTTAA 2196
||||||||||||||||||||||||||||||||||||
Db 2161 ATGTACAGCAGGGCATATGAAACACATACGCTTTAA 2196