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 .
Response to Previous Communication
In the remarks filed on 05/14/2026 and made in response to restriction on 02/24/2026, Applicant elects Group 1 for prosecution on the merits. Group 1 is directed to Claims 1, 3, 7, 10, 14-15, and 47 encompassing a transgenic plant comprising a synthetic polynucleotide expression cassette encoding the animal-derived protein, myoglobin. Additionally, Applicant provisionally elects (i) Nucleic Acid sequences with SEQ ID NO: 78; (ii) SEQ ID NO: 65; and (iii) Amino Acid sequences with SEQ ID NO: 58 for prosecution on the merits with traversal. Applicant further alleges that Claims 1, 3, 7, 10, 14, 15 and 47 are generic and/or read on the provisionally elected species and submits that there would be no undue burden to search all the identified Species. Applicant argues that the election of species should be withdrawn. Because Applicants did not distinctly and specifically point out supposed errors in the restriction requirement with respect to groups of inventions, the election has been treated as an election without traverse. See MPEP § 818.03(a).
Regarding search burden (i.e., traversal with respect to species election), Applicants are reminded that the assertion of a search burden in a lack of unity of invention practice under PCT Rule 13.1 and 13.2 is irrelevant, therefore demonstration of a search burden is not required.
Status of Claims
Claims 1,3,7,10,14-15,32-34,38-43, 47, 59, 65, 70-71, and 83 are pending.
Claims 32-34,38-43, 59, 65, 70-71, and 83 are withdrawn under the restriction requirement as being directed to non-elected invention(s).
Claims 1,3,7,10,14-15, and 47 are examined herein.
Claim Objections
Claim 1 is objected to for adding an extra space between myoglobin and the comma in the third line.
Claim 14 is objected to for adding an unnecessary comma after “32” in the fourth line.
Information Disclosure Statement
Foreign patent documents 2 and 3 listed in IDS filed on 05/14/2026 had no English translation other than the abstract. The listing of references on pages 72-74 of the specification is not a proper information disclosure statement. 37 CFR 1.98(b) requires a list of all patents, publications, or other information submitted for consideration by the Office, and MPEP § 609.04(a) states, "the list may not be incorporated into the specification but must be submitted in a separate paper." Therefore, unless the references have been cited by the examiner on form PTO-892 or in IDS documents filed on 05/14/2026 or 02/07/2025, they have not been considered.
Specification
The disclosure is objected to because it contains an embedded hyperlink and/or other form of browser-executable code (see paragraphs 131 and 134 for examples). Applicant is required to delete the embedded hyperlink and/or other form of browser-executable code; references to websites should be limited to the top-level domain name without any prefix such as http:// or other browser-executable code. See MPEP § 608.01.
Examples 5-7 appear to be missing.
Definitions referenced in the specification such as “AxB” maize in paragraph 163 and “T0” in paragraph 208 appear undefined. Applicant is encouraged to read the specification and ensure all terms are defined.
Claim Interpretations
Claim 14 is directed to a DNA sequence (SEQ ID NO: 78) which is identical to the codon optimized sequence (SEQ ID NO:32 in Table 3) with the exception of a nucleic acid change from ATC to CAC changing the amino acid ILE to a HIS and was described in Paragraph 206 of the Specification. Therefore, SEQ ID NO: 78 is interpreted to be codon optimized SEQ ID NO:32.
Claim 47 is directed to myoglobin protein included in the expression cassette that comprises a sequence with at least 90% identity to a reference sequence SEQ ID NO: 65. The expression cassette’s components are described in table 5 on page 51 and table 6 page 52. The examiner interprets the construct to encode:
1) Rice glutelin B-4 promoter (SEQ ID NO: 88)
2) Maize gamma zein 27 signal peptide (SEQ ID NO: 24)
3) MYGm (i.e. myoglobin) coding sequence (SEQ ID NO: 78)
4) “T35S” terminator
Improper Markush Group
Claims 15 and 47 are rejected under the judicially-created basis that they contain an improper Markush groupings of alternative species. See In re Harnisch, 631 F.2d 716, 721-722 (CCPA 1980) and Ex parte Hozumi, 3 USPQ2d 1059, 1060 (Bd. Pat. App. and Int. 1984). The improper Markush groupings includes species of the claimed invention that do not share both a substantial structural feature and a common use that flows from the substantial structural feature. All dependent claims are included in these rejections unless they contain a limitation that overcomes the deficiencies of the parent claim from which they depend.
The members of the improper Markush groupings do not share a substantial feature and a common use that flows from the substantial structural feature for the following reasons: the claims encompass thousands of unique polypeptide sequences (and the corresponding thousands of unique nucleic acid sequences), which are biochemically divergent, they have no conserved structure throughout the genus other than a phosphodiester backbone or a polypeptide backbone (peptide linkages), respectively, and may or may not be associated with synthesizing myoglobin in monocotyledonous plant’s grain.
The species recited in Claim 15 lack a substantial structural feature for the claimed 95% amino acid similarity as evidenced by the failure of the non-elected sequence (SEQ ID NO: 3) to claim a sequence with similarity higher than the claimed 95% similarity to the elected sequence, SEQ ID NO:58. See alignment between SEQ ID NO: 58 and SEQ ID NO: 3 below, demonstrating only 91% sequence identity.
Score
Expect
Method
Identities
Positives
Gaps
273 bits(697)
1e-100
Compositional matrix adjust.
135/154(88%)
141/154(91%)
0/154(0%)
Query 1 MGLSDGEWQLVLNAWGKVEADVAGHGQEVLIRLFTGHPETLEKFDKFKHLKTEAEMKASE 60
MGLSDGEWQLVLN WGKVEADVAGHGQEVLIRLF GHPETLEKFDKFKHLK+E EMKASE
Sbjct 1 MGLSDGEWQLVLNVWGKVEADVAGHGQEVLIRLFKGHPETLEKFDKFKHLKSEDEMKASE 60
Query 61 DLKKHGNTVLTALGGILKKKGHHEAEVKHLAESHANKHKIPVKYLEFHSDAIIHVLHAKH 120
DLKKHGNTVLTALGGILKKKGHHEAE+ LA+SHA KHKIPVKYLEF S+AII VL +KH
Sbjct 61 DLKKHGNTVLTALGGILKKKGHHEAELTPLAQSHATKHKIPVKYLEFISEAIIQVLQSKH 120
Query 121 PSDFGADAQAAMSKALELFRNDMAAQYKVLGFHG 154
P DFGADAQ AMSKALELFRNDMAA+YK LGF G
Sbjct 121 PGDFGADAQGAMSKALELFRNDMAAKYKELGFQG 154
QUERY is SEQ ID NO: 58
SUBJECT is SEQ ID NO: 3
The species recited in Claim 47 lack a substantial structural feature for the claimed 95% nucleic acid similarity as evidenced by the failure of the non-elected sequence (SEQ ID NO: 48) to claim a sequence with a sequence similarity higher than 95% similarity to the elected sequence of SEQ ID NO:65. A NCBI alignment between SEQ ID NO: 48 and SEQ ID NO: 65 only align in 24% of the query sequence (i.e. SEQ ID NO: 65) while there is no detectable similarity between the remaining 76% (see NCBI alignment result below).
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625
1273
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Greyscale
Thus, the species of each of Claims 15 and 47 do not share both a substantial structural feature and a common use which flows therefrom.
In response to this rejection, Applicants should either amend the claim(s) to recite only individual species or grouping of species that share a substantial structural feature as well as a common use that flows from the substantial structural feature, or present a sufficient showing that the species recited in the alternative of the claims(s) in fact share a substantial structural feature as well as a common use that flows from the substantial structural feature.
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.
Written Description
Claims 1,3,7, and 10 are rejected under 35 USC § 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 applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention.
The Federal Circuit has clarified the written description requirement. The court stated that a written description of an invention "requires a precise definition, such as by structure, formula, [or] chemical name, of the claimed subject matter sufficient to distinguish it from other materials". University of California v. Eli Lilly and Co., 119 F.3d 1559, 1568; 43 USPQ2d 1398, 1406 (Fed. Cir. 1997). The court also concluded that "naming a type of material generally known to exist, in the absence of knowledge as to what that material consists of, is not description of that material". Id. Further, the court held that to adequately describe a claimed genus, Patent Owner must describe a representative number of the species of the claimed genus, and that one of skill in the art should be able to "visualize or recognize the identity of the members of the genus".
The claims are broadly drawn to a transgenic monocotyledonous plant comprising a synthetic polynucleotide encoding at least one animal-derived protein (i.e. elected myoglobin). The plant’s grain expresses the animal-derived protein at a level in the range from 0.01 mg to 24.0 mg of the animal-derived protein per gram of grain.
Applicant describes:
"Myoglobin" is defined as heme-containing globular protein found in the cardiac and skeletal muscle tissue of vertebrates in general and in almost all mammals in paragraph 43. Bovine, chicken, and swine, myoglobin polypeptide sequences (i.e. SEQ ID NOs: 1-3) and a truncated Bovine myoglobin (SEQ ID NO: 20) with a targeting sequences (SEQ ID NO: 40) are also described. SEQ ID NO: 58 is also described, which is a mutated version of myoglobin which is described in paragraph 206 as having a strong bond with heme resulting in a protein that is less susceptible to degradation. Described myoglobin polynucleotide sequences were SEQ ID NOs: 32 which encodes the polypeptide sequence of SEQ ID NO:1 while the polynucleotide sequence SEQ ID NO: 78 encodes the mutated myoglobin polypeptide sequence SEQ ID NO:58. These sequences were codon optimized.
Paragraph 194 teaches a plasmid pAG5025 (SEQ ID NO:65 of Claim 47 and Figure 4B) as well as the maize ferrochelatase FC1A were expressed in the maize seed under the rice glutelin-1 gene or the maize gamma zein 27 kDa gene promoters.
Paragraph 162 of example 2 teach pAG5026, pAG5016, pAG5019, and pAG5020 were used to transform embryogenic callus from wild type maize. These vector contains the unmutated myoglobin sequence. The vector pAG5026 encoded the mutated and unmutated version of myoglobin (see tables 5 and 6) .
Figures 7 shows that plants transformed with these vectors were able to express myoglobin in the seeds while figure 8 shows myoglobin was detected in seeds from plants transformed with pAG5026.
Applicant does not describe:
Description of any other myoglobin besides those of polypeptides of SEQ ID NOs: 1-3,20, 40, and 58 and polynucleotide sequences of SEQ ID NOs: 32 and 78.
Protein expression results from plants transformed with pAG5025
Transformation of any other vector containing a myoglobin polypeptide sequence other than those outlined in SEQ ID NOs: 1 and 58 (encoded by polynucleotide sequences of SEQ ID NOs: 32 and 78), or with any regulatory elements other than Rice Glutelin-1 Rice or Glutelin B-4 promoter, maize gamma zein 27 signal peptides, and NOS or T35S terminators.
Expressing a myoglobin protein in the grain of any other monocotyledonous plant.
The instant disclosure and the instant claims do not set forth clear structural characteristics essential to identifying myoglobin being expressed in a transgenic monocotyledonous plant. For example, other proteins meet the myoglobin definition outlined by the specification. Cytoglobin proteins are heme-containing globular protein and are described in page 15 paragraph 6 of Vlasova, Irina I. "Peroxidase activity of human hemoproteins: keeping the fire under control." Molecules 23.10 (2018): 2561. Cytoglobin is also found in mammal muscle cells as recited by Singh, Sarvjeet, et al. "Cytoglobin modulates myogenic progenitor cell viability and muscle regeneration." Proceedings of the National Academy of Sciences 111.1 (2014): E129-E138 in the passage “Cygb [i.e. cytoglobin] is a heme-containing protein of the globin family that is structurally similar to myoglobin (Mb) and is conserved across species (mammals, fish, amphibians, reptiles, and birds). In mammals it is expressed in a variety of tissues, including skeletal muscle”. Because multiple proteins fit the myoglobin definition provided in the specification, it would be difficult to one of ordinary skill in the art to identify if a protein is encompassed by the limitations of Claims 1,3,7, and 10.
The Applicant has provided only the working example for the protein with polypeptide sequences of SEQ ID NOs: 1 and 58 in maize kernels and has a broad definition of myoglobin. Therefore, given the lack of written description in the instant disclosure with regard to the structural and functional characteristics of the claimed compositions, Applicant does not appear to have been in possession of the claimed genus at the time this application was filed.
Scope of Enablement
Claims 1,3,15, and 47 are rejected under 35 USC § U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, because the specification, while being enabling for expressing myoglobin with specific SEQ ID NOs in maize kernels, does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make or use the invention commensurate in scope with these claims.
In re Wands lists a number of factors for determining whether or not undue experimentation would be required by one skilled in the art to make and/or use the invention. These factors are: (1) the quantity of experimentation necessary; (2) the amount of direction or guidance presented; (3) the presence or absence of working examples of the invention; (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; (8) the breadth of the claim. In re Wands, 858 F.2d 731, 8 USPQ2d 1400 (Fed. Cir. 1988).
Claims 1,3,15, and 47 are broadly directed to transgenic monocotyledonous plant or tissue thereof comprising a synthetic polynucleotide encoding expressing an animal-derived myoglobin protein in the range from 0.01 mg to 24.0 mg of the animal-derived protein per gram of grain.
Applicant Teaches:
"Myoglobin" is defined as heme-containing globular protein found in the cardiac and skeletal muscle tissue of vertebrates in general and in almost all mammals in paragraph 43. Bovine, chicken, and swine, myoglobin polypeptide sequences (i.e. SEQ ID NOs: 1-3) and a truncated Bovine myoglobin (SEQ ID NO: 20) with a targeting sequences (SEQ ID NO: 40) are also described. SEQ ID NO: 58 is also described, which is a mutated version of myoglobin which is described in paragraph 206 as having a strong bond with heme resulting in a protein that is less susceptible to degradation. Provided myoglobin polynucleotide sequences were SEQ ID NOs: 32 which encodes the polypeptide sequence of SEQ ID NO:1 and the polynucleotide sequence SEQ ID NO: 78 which encodes the mutated myoglobin polypeptide sequence SEQ ID NO:58. These sequences were codon optimized.
Paragraph 194 provides a plasmid pAG5025 (SEQ ID NO:65 of Claim 47 and Figure 4B) as well as the maize ferrochelatase FC1A were expressed in the maize seed under the rice glutelin-1 gene or the maize gamma zein 27 kDa gene promoters.
Paragraph 162 of example 2 provides pAG5026, pAG5016, pAG5019, and pAG5020 which were used to transform embryogenic callus from wild type maize. These vector contains the unmutated myoglobin sequence. The vector pAG5026 encoded the mutated and unmutated version of myoglobin (see tables 5 and 6) .
Figures 7 shows that plants transformed with these vectors were able to express myoglobin in the seeds while figure 8 shows myoglobin was detected in seeds from plants transformed with pAG5026.
Applicant Does Not Teach:
Applicant does not provide any other myoglobin polypeptides besides those of SEQ ID NOs: 1-3,20, 40, and 58.
Protein expression results from plants transformed with pAG5025
Applicant does not provide an example of a transformation event using any other vector containing a myoglobin polypeptide sequence other than those outlined in SEQ ID NOs: 1 and 58 (encoded by polynucleotide sequences of SEQ ID NOs: 32 and 78), or with any regulatory elements other than Rice Glutelin-1 Rice or Glutelin B-4 promoter, maize gamma zein 27 signal peptides, and NOS or T35S terminators.
Applicant does not provide any example of a transgenic monocotyledonous plant expressing a myoglobin protein in the grain.
The instant disclosure and the instant claims do not set forth structural characteristics essential to expressing myoglobin in all monocotyledonous species’ grain at a level between 0.01 to 24 mg of the animal-derived protein per gram of grain as the claims are directed to.
It is known in the art that some common monocotyledonous plants are recalcitrant to transformation. For example, Claim 3 is directed to expressing myoglobin in wheat grain. However, the abstract of Guo, Guanghui, et al. "An elegant co‐transformation strategy for recalcitrant wheat using morphogenic regulators." The Plant Journal 124.2 (2025): e70541 teaches wheat transformation efficiency largely depends on the recipient genotype and overcoming low regeneration efficiency is a barrier to successfully transforming wheat. Morphogenic regulators are shown to improve wheat transformation and regeneration, however, there are still issues regarding how to specifically utilize them in recalcitrant wheat. These challenges are described on page 2 in paragraphs 1-2 and solutions for overcoming these barriers were not provided in the disclosure.
The Applicant has provided only the working example for expressing myoglobin with polypeptide sequences of SEQ ID NOs:1 and 58 (encoded by polynucleotide sequences of SEQ ID NOs: 32 and 78) in maize seeds, but not in any other plant species. Thus, the examples provided by the Applicant do not provide adequate working examples to enable the scope of the invention without undue experimentation. Given the breadth of the claims, the lack of guidance and working examples, the unpredictability in the art, and the state of the art, undue experimentation would be required to make and use the claimed invention, and therefore, the invention is not enabled throughout the broad scope of the claims.
Claim Rejections - 35 USC § 102/103
A rejection under 35 U.S.C. § 102 or § 103 does not require the same analysis as a rejection under 35 U.S.C. § 102 or a rejection under 35 U.S.C. § 103. The rejection is made because the examiner cannot determine whether the prior art composition possesses characteristics that are not recited in the art. The examiner does not have sufficient facts to determine whether the claimed compositions, polynucleotides, polypeptides and organisms are inherently the same as the prior art compositions, polynucleotides, polypeptides and organisms. In addition, the examiner cannot conclude that the claimed subject matter would have been obvious since it cannot be determined whether the claimed and prior art compositions, polynucleotides, polypeptides and organisms differ. Where the prior art product seems to be identical, except that the prior art is silent to a characteristic or property claimed, then the burden shifts to applicants to provide evidence that the prior art would neither anticipate nor render obvious the claimed invention. In re Best, 195 USPQ 430, 433 (CCPA 1977).
Claims 1,3,7, 10, and 15 are rejected as being anticipated under 35 U.S.C. § 102(a)(1), or alternatively as being unpatentable under 35 U.S.C. § 103, over US 2016/0340411 Al.
Claims 1,3,7, 10, and 15 are directed to a transgenic monocotyledonous plant or tissue thereof comprising a synthetic polynucleotide encoding at least one animal-derived myoglobin, hemoglobin, chymosin, or casein protein. The plant’s grain expresses the animal-derived protein at a level in the range from 0.01 mg to 24.0 mg of the animal-derived protein per gram of grain. The monocotyledonous plant is ideally corn and the animal-derived protein is myoglobin. The myoglobin has a polypeptide sequence at least 95% identical to SEQ ID NO: 58.
US 2016/0340411 Al relates to methods and material for producing heme-containing polypeptides (i.e. myoglobin or hemoglobin), and more particularly, to producing heme-containing polypeptides in recombinant bacterial cells such as Bacillus cells or in recombinant plants or plant cells. Paragraph 7 teaches a recombinant plant, such as Zea mays (i.e. Claims 1,3 and 7), producing a heme containing polypeptide. This paragraph also teaches the exogenous nucleic acid can include a regulatory control element such as tissue specific promoters for seeds (i.e. Claim 1). The heme-containing polypeptide can be a myoglobin, which is taught in paragraph 14 while paragraph 44 teaches the myoglobin from Bos Taurus (SEQ ID NO: 18 in Figure 9) which has 99.4% identity (see alignment below) to applicant’s SEQ ID NO: 58 (Claim 1,10, and 15). These limitations are further reiterated in Claims 19,23,26,27-29 of US 2016/0340411 Al.
99.4% identity in 154 residues overlap; Score: 800.0; Gap frequency: 0.0%
Sequence1 1 MGLSDGEWQLVLNAWGKVEADVAGHGQEVLIRLFTGHPETLEKFDKFKHLKTEAEMKASE
Sequence2 1 MGLSDGEWQLVLNAWGKVEADVAGHGQEVLIRLFTGHPETLEKFDKFKHLKTEAEMKASE
************************************************************
Sequence1 61 DLKKHGNTVLTALGGILKKKGHHEAEVKHLAESHANKHKIPVKYLEFISDAIIHVLHAKH
Sequence2 61 DLKKHGNTVLTALGGILKKKGHHEAEVKHLAESHANKHKIPVKYLEFHSDAIIHVLHAKH
*********************************************** ************
Sequence1 121 PSDFGADAQAAMSKALELFRNDMAAQYKVLGFHG
Sequence2 121 PSDFGADAQAAMSKALELFRNDMAAQYKVLGFHG
**********************************
The property of expressing animal-derived myoglobin in grain in the range from 0.01 mg to 24.0 mg of the animal-derived protein per gram of grain would be inherent to the compositions taught by US 2016/0340411 Al (polypeptide that is 99.4% similar to instant SEQ ID NO:58, transgenic plants, and tissue specific promoters), which teaches the recited structure(s) that are required for the claimed function(s).
Accordingly, US 2016/0340411 Al anticipated the claimed invention.
Alternatively, it is possible that US 2016/0340411 Al does not explicitly teach every single element of the instant claims, and perhaps this could result in compositions that are not identical to the instantly claimed composition(s). However, it would have been prima facie obvious and within the scope of an ordinary skill in the art at the time the application was filed to use the compositions and methods taught by US 2016/0340411 Al, to check and select seeds with the protein (i.e. myoglobin or heme-containing polypeptide) content for expression level in the range from 0.01 mg to 24.0 mg of the animal-derived protein per gram of grain, and this would result in the Applicant’s invention; with a reasonable expectation of success, and without any surprising results. One of ordinary skill in the art would have known to check and select seeds with the protein (i.e. myoglobin or heme-containing polypeptide) content for expression level in the range from 0.01 mg to 24.0 mg of the animal-derived protein per gram of grain.“providing a consumable that can be substantially or entirely composed of ingredients derived from non-animal sources, yet recapitulates key features associated with the cooking and consumption of an equivalent meat product derived from animals” which would meet the “need for methods to produce proteins at large scale for industrial and food purposes” and was described in paragraphs 3 and 196 of US 2016/0340411 Al.
Claim Rejections - 35 USC § 103
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 (i.e., changing from AIA to pre-AIA ) 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 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.
Claims 14 and 47 are rejected under 35 U.S.C. 103 as being unpatentable over US 2016/0340411 Al as they are applied to Claim 1 above in regards to the myoglobin protein and expressing animal-derived protein in the grain of a monocot plant (i.e. corn), and further in view of US 20130071884 A1 and US 20030145347 A1.
Claims 14 and 47 teach a myoglobin protein encoded by the nucleic acid with at least 90% identity to codon optimized SEQ ID NO:79. The claims also encompass an expression cassette that comprises a sequence with at least 90% identity to a reference sequence SEQ ID NOs: 65. The expression cassette’s components are described in table 5 on page 51 and table 6 page 52 of the specification. The construct encodes:
1) Rice glutelin B-4 promoter position 1-1474 (SEQ ID NO: 88)
2) Maize gamma zein 27 signal peptide position 1484-1540 (SEQ ID NO: 24 )
3) MYGm (i.e. myoglobin) coding sequence position 1541-2014 (SEQ ID NO 78)
4) “T35S” terminator position 2021-2296
US 2016/0340411Al relates to methods and material for producing heme-containing polypeptides (i.e. myoglobin or hemoglobin) in recombinant plants or plant cells as was described previously. Paragraph 44 teaches the myoglobin from Bos Taurus (SEQ ID NO: 18 in Figure 9) which has 99.4% identity to applicant’s SEQ ID NO: 58 which is the translated product from SEQ ID NO: 78 (The examiner notes the polynucleotide lacks the start codon) of Claim 14. Paragraph 70 teaches the myoglobin sequence can be codon optimized while paragraph 73 teaches codon optimization can be performed for optimizing gene expression in plants (i.e. Codon optimized gene in Claim 14).
US 2016/0340411Al does not explicitly teach the use of an expression cassette that comprises a sequence with at least 90% identity to a reference sequence SEQ ID NOs: 65.
US 20130071884 A1 is directed to vectors for expression of proteins in plants and appear to be disclose by the same applicant and inventors. The vectors in this application are also described as being able to express genes or enzymes, besides the cell wall degrading enzymes the application is directed to, and is stated in paragraph 92 and the abstract while paragraph 93 states these vectors can be used to transform monocotyledonous plants. One disclosed vector, SEQ ID NO: 274, bears resemblance to the presently claimed construct and is described in paragraph 219. Paragraph 219 describes this sequence contains a Rice Glutelin B-4 promoter and a “Nos” terminator. The Applicant’s construct sequence position 1-1474 (underlined in Query sequence below) aligns to this construct and SEQ ID NO: 4 of US 20130071884 A1 indicating that this Glutelin B-4 promoter sequence is known in the art. Paragraph 112 teaches this sequence is a seed specific promoter. The termination region of SEQ ID NO: 65 also aligns to this construct, but the termination sequence is called a “Nos” terminator. Nos terminators are described in paragraph 118, which also teaches 35S terminator can be used to replace them, and the sequence given (SEQ ID NO: 29) align to the termination region in Applicant’s SEQ ID NO 65 (bolded in Query sequence below).
%
Result Query Filing
No. Score Match Length ID Date Dups Description
-------------------------------------------------------------------------------------------------------------
18 1568.6 68.3 12400 US-13-508-280A-274 2012-09-20 2 PLANTS EXPRESSING CELL WALL DEGRADING ENZYMES AND EXPRESSION VECTORS
ALIGNMENT:
Query Match 68.3%; Score 1568.6; Length 12400;
Best Local Similarity 80.4%;
Matches 1970; Conservative 0; Mismatches 324; Indels 157; Gaps 5;
Qy 1 TACAGGGTTCCTTGCGTGAAGAAGGGTGGCCTGCGGTTCACCATTAACGGTCACGACTAC 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 9951 TACAGGGTTCCTTGCGTGAAGAAGGGTGGCCTGCGGTTCACCATTAACGGTCACGACTAC 10010
Qy 61 TTCCAGCTAGTACTGGTGACCAACGTCGCGGCGGCAGGGTCAATCAAGTCCATGGAGGTT 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 10011 TTCCAGCTAGTACTGGTGACCAACGTCGCGGCGGCAGGGTCAATCAAGTCCATGGAGGTT 10070
Qy 121 ATGGGTTCCAACACAGCGGATTGGATGCCGATGGCACGTAACTGGGGCGCCCAATGGCAC 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 10071 ATGGGTTCCAACACAGCGGATTGGATGCCGATGGCACGTAACTGGGGCGCCCAATGGCAC 10130
Qy 181 TCACTGGCCTACCTCACCGGTCAAGGTCTATCCTTTAGGGTCACCAACACAGATGACCAA 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 10131 TCACTGGCCTACCTCACCGGTCAAGGTCTATCCTTTAGGGTCACCAACACAGATGACCAA 10190
Qy 241 ACGCTCGTCTTCACCAACGTCGTGCCACCAGGATGGAAGTTTGGCCAGACATTTGCAAGC 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 10191 ACGCTCGTCTTCACCAACGTCGTGCCACCAGGATGGAAGTTTGGCCAGACATTTGCAAGC 10250
Qy 301 AAGCTGCAGTTCAAGTGAGAGGAGAAGCCTGAATTGATACCGGAGCGTTTCTTTTGGGAG 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 10251 AAGCTGCAGTTCAAGTGAGAGGAGAAGCCTGAATTGATACCGGAGCGTTTCTTTTGGGAG 10310
Qy 361 TAACATCTCTGGTTGCCTAGCAAACATATGATTGTATATAAGTTTCGTTGTGCGTTTATT 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 10311 TAACATCTCTGGTTGCCTAGCAAACATATGATTGTATATAAGTTTCGTTGTGCGTTTATT 10370
Qy 421 CTTTCGGTGTGTAAAATAACATACATGCTTTCCTGATATTTTCTTGTATATATGTACACA 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 10371 CTTTCGGTGTGTAAAATAACATACATGCTTTCCTGATATTTTCTTGTATATATGTACACA 10430
Qy 481 CACACGACAAATCCTTCCATTTCTATTATTATTGAACAATTTAATTGCGAGGGCGAGTAC 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 10431 CACACGACAAATCCTTCCATTTCTATTATTATTGAACAATTTAATTGCGAGGGCGAGTAC 10490
Qy 541 TTGTCTGTTTACCTTTTTTTTTTCAGATGGCATTTTATAGTTTAACCTTTCATGGACCGG 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 10491 TTGTCTGTTTACCTTTTTTTTTTCAGATGGCATTTTATAGTTTAACCTTTCATGGACCGG 10550
Qy 601 CAGTAGTTCTAACCATGAATGAAAAGAAATCATAGTCCACACCACGCAGGGACATTGTGG 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 10551 CAGTAGTTCTAACCATGAATGAAAAGAAATCATAGTCCACACCACGCAGGGACATTGTGG 10610
Qy 661 TCATTTTAGACAAGACGATTTGATTAATGTCTTGTATGATATGGTCGACAGTGAGGACTA 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 10611 TCATTTTAGACAAGACGATTTGATTAATGTCTTGTATGATATGGTCGACAGTGAGGACTA 10670
Qy 721 ACAAACATATGGCATATTTTATTACCGGCGAGTTAAATAAATTTATGTCACAGTAATAAA 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 10671 ACAAACATATGGCATATTTTATTACCGGCGAGTTAAATAAATTTATGTCACAGTAATAAA 10730
Qy 781 CTGCCTAATAAATGCACGCCAGAAAATATAATGATAAAAAAAAGAAAAGATACATAAGTC 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 10731 CTGCCTAATAAATGCACGCCAGAAAATATAATGATAAAAAAAAGAAAAGATACATAAGTC 10790
Qy 841 CATTGCTTCTACTTTTTTAAAAATTAAATCCAACATTTTCTATTTTTTGGTATAAACTTG 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 10791 CATTGCTTCTACTTTTTTAAAAATTAAATCCAACATTTTCTATTTTTTGGTATAAACTTG 10850
Qy 901 GAAGTACTAGTTGGATATGCAAAATCATCTAACCTCCATATATTTCATCAATTTGTTTAC 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 10851 GAAGTACTAGTTGGATATGCAAAATCATCTAACCTCCATATATTTCATCAATTTGTTTAC 10910
Qy 961 TTTACATATGGGAGAGGATAGTATGTCAAAGAAAATGACAACAAGCTTACAAGTTTCTTA 1020
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 10911 TTTACATATGGGAGAGGATAGTATGTCAAAGAAAATGACAACAAGCTTACAAGTTTCTTA 10970
Qy 1021 TTTTAAAAGTTCCGCTAACTTATCAAGCATAGTGTGCCACGCAAAACTGACAACAAACCA 1080
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 10971 TTTTAAAAGTTCCGCTAACTTATCAAGCATAGTGTGCCACGCAAAACTGACAACAAACCA 11030
Qy 1081 ACAAATTTAAGGAGCGCCTAACTTATCATCTATGACATACCGCACAAAATGATAACATAC 1140
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 11031 ACAAATTTAAGGAGCGCCTAACTTATCATCTATGACATACCGCACAAAATGATAACATAC 11090
Qy 1141 TAGAGAAACTTTATTGCACAAAAGGAAATTTATCCATAAGGCAAAGGAACATCTTAAGGC 1200
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 11091 TAGAGAAACTTTATTGCACAAAAGGAAATTTATCCATAAGGCAAAGGAACATCTTAAGGC 11150
Qy 1201 TTTGGATATACATTTACCAACAAGCATTGTTTGTATTACCCCTAAAGCGCAAGACATGTC 1260
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 11151 TTTGGATATACATTTACCAACAAGCATTGTTTGTATTACCCCTAAAGCGCAAGACATGTC 11210
Qy 1261 ATCCATGAGTCATAGTGTGTATATCTCAACATTGCAAAGCTACCTTTTTTCTATTATACT 1320
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 11211 ATCCATGAGTCATAGTGTGTATATCTCAACATTGCAAAGCTACCTTTTTTCTATTATACT 11270
Qy 1321 TTTCGCATTATAGGCTAGATATTATCTATACATGTCAACAAACTCTATCCCTACGTCATA 1380
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 11271 TTTCGCATTATAGGCTAGATATTATCTATACATGTCAACAAACTCTATCCCTACGTCATA 11330
Qy 1381 TCTGAAGATTCTTTTCTTCACTATATAAGTTGGCTTCCCTGTCATTGAACTCACATCAAC 1440
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 11331 TCTGAAGATTCTTTTCTTCACTATATAAGTTGGCTTCCCTGTCATTGAACTCACATCAAC 11390
Qy 1441 CAGCCCAAGTTTCCAATAACATCCTCAAATAGCTGGATCCACCATGAGGGTGTTGCTCGT 1500
|||||||||||||||||||||||||||||||||| ||||||| | | | | |
Db 11391 CAGCCCAAGTTTCCAATAACATCCTCAAATAGCTATGGCCACCATCGCTTTCTCCCGCTT 11450
Qy 1501 TGCCCTCGCTCT---CCTGGCTCTCGCTGCGAGCGCCACCAGCGGGTTAAGTGATGGAGA 1557
|| || | | || |||| || | ||| | || | |||| |
Db 11451 GTCCATCTACTTCTGCGTGCTTCTCCTGTGCCACGGCTCCATGGCCTTCCCAGCTGGAAA 11510
Qy 1558 ATGGCAGCTGGTGCTCAATGCATGGGGCAAGGTGGAGGCAGATGTGGCGGGTCACGGCCA 1617
| || || | || | | || | | | |||| |
Db 11511 CGCAACGGAATTGGAGAAAAGACAAACCACCCCTAACTCTGAGGGCTGGCATGACGGATA 11570
Qy 1618 AGAAGTTCTCATCAGGCTCTTTACCGGTCATCCAGAGACGCTGGAGAAGTTTGACAAGTT 1677
| || | | ||| | | || || | ||
Db 11571 CTACTACTCTTGGTGGAGCGATGGTGGTGCACAGGCCACCTATACAAACCTCGAAGGCGG 11630
Qy 1678 CAAGCATCTGAAAACAGAAGCAGAGATGAAGGCCAGCGAGGATCTGAAGAAACACGGTAA 1737
|| | | ||| | | | | || | ||| | | | | | | || |
Db 11631 CA--CTTATGAGATTTCATGGGGTGACGGTGGCAACCTTGTCGGCGGAAAGGGGTGGAAC 11688
Qy 1738 CACTGTCCTTACAGCTCTGGGCGGCATTTTGAAGAAG-----------AAGGGACACCAT 1786
| | | | | || | | || || |||| | || || ||
Db 11689 CCCGGACTTAACGCCAGGGCAATCCACTTCGAAGGGGTGTACCAGCCCAATGGCAACTCA 11748
Qy 1787 GAAGCCGAGGTCAAACATTTGGCAGAATCTCACGCCAACAAGCACAAGATACCGGTAAAA 1846
| | ||| | | | | || || || | || || ||
Db 11749 TACCTGGCCGTCTACGGGTGGACGCGCAATCCGCTGGTTGAGTACTATATCGTGGAGAAT 11808
Qy 1847 TATCTAGAGTTCCAC---AGCGACGCCATCATTCACGTCCTGCACGCCAAGCACCCTTCA 1903
| | | || ||| || || || || | | | || |
Db 11809 TTCGGAACTTATGACCCTAGCTCCGGTGCCACGGACCTCGGGACAGTCGAGTGTGACGGA 11868
Qy 1904 GACTTCGGCGCTGACGCTCAAGCGGCCATGTCCAAAGCGCTGGAGCTGTTCCGCAATGAT 1963
| || | | | || | || || || | | | | ||| |
Db 11869 AGCATCTACAGGCTGGGTAAAACTACCCGCGTTAATGCTCCATCGATCGACGGCACGCAA 11928
Qy 1964 ATGGCCGCGCAGTACAAGGTTCTTGGGTTTCATGGAAAG--------------------- 2002
| | || ||| | | || || | |
Db 11929 ACATTTGATCAATACTGGTCCGTGCGGCAGGATAAGAGGACAAGCGGCACAGTTCAGACG 11988
Qy 2003 ------------------------------------------------------------ 2002
Db 11989 GGTTGCCACTTTGATGCCTGGGCAAGAGCGGGGCTCAATGTGAATGGGGACCACTACTAT 12048
Qy 2003 ---------------------------------------------------------GAC 2005
||
Db 12049 CAGATTGTGGCGACCGAGGGCTATTTCTCCAGTGGCTATGCGCGTATAACCGTCGCTGAT 12108
Qy 2006 GAGCTGTGACCTAGGTCCCCGAATTTCCCCGATCGTTCAAACATTTGGCAATAAAGTTTC 2065
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 12109 GTTGGATGACCTAGGTCCCCGAATTTCCCCGATCGTTCAAACATTTGGCAATAAAGTTTC 12168
Qy 2066 TTAAGATTGAATCCTGTTGCCGGTCTTGCGATGATTATCATATAATTTCTGTTGAATTAC 2125
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 12169 TTAAGATTGAATCCTGTTGCCGGTCTTGCGATGATTATCATATAATTTCTGTTGAATTAC 12228
Qy 2126 GTTAAGCATGTAATAATTAACATGTAATGCATGACGTTATTTATGAGATGGGTTTTTATG 2185
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 12229 GTTAAGCATGTAATAATTAACATGTAATGCATGACGTTATTTATGAGATGGGTTTTTATG 12288
Qy 2186 ATTAGAGTCCCGCAATTATACATTTAATACGCGATAGAAAACAAAATATAGCGCGCAAAC 2245
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 12289 ATTAGAGTCCCGCAATTATACATTTAATACGCGATAGAAAACAAAATATAGCGCGCAAAC 12348
Qy 2246 TAGGATAAATTATCGCGCGCGGTGTCATCTATGTTACTAGATCGGGAATTG 2296
|||||||||||||||||||||||||||||||||||||||||||||||||||
Db 12349 TAGGATAAATTATCGCGCGCGGTGTCATCTATGTTACTAGATCGGGAATTG 12399
US 20130071884 A1 does not explicitly teach the region of the construct that encodes the maize gamma zein 27 signal peptide position 1484-1540 (SEQ ID NO: 24).
US 20030145347 A1 is a patent publication directed to improving methods of grain processing to enhance protein and starch recovery, particularly in corn wet milling (see paragraph 1). The maize gamma zein 27 signal peptide is taught in US 20030145347 A1 in paragraph 253 where primers (SEQ ID NOs: 14-15) and have 100% identity to the Applicant’s maize gamma zein 27 signal peptide (See alignment below). It is taught, in conjunction with a gamma zein promoter, to target protein to the endomembrane system of endosperm cells (see paragraphs 246 and 253). Vector using this sequences were used to transform maize (paragraph 254).
US-10-306-292-14
Sequence 14, US/10306292
Publication No. US20030145347A1
GENERAL INFORMATION
APPLICANT: Lanahan, Michael B.
APPLICANT: Desai, Nalini M.
APPLICANT: Gasdaska, Pamela Y.
TITLE OF INVENTION: GRAIN PROCESSING METHOD AND TRANSGENIC PLANTS USEFUL
TITLE OF INVENTION: THEREIN
FILE REFERENCE: A-31383P1
CURRENT APPLICATION NUMBER: US/10/306,292
CURRENT FILING DATE: 2002-11-27
PRIOR APPLICATION NUMBER: US/09/598,747
PRIOR FILING DATE: 2000-06-21
NUMBER OF SEQ ID NOS: 42
SEQ ID NO 14
LENGTH: 65
TYPE: DNA
ORGANISM: Artificial Sequence
FEATURE:
OTHER INFORMATION: Description of Artificial Sequence:
oligonucleotide (primer NMD126)
Query Match 100.0%; Score 57; Length 65;
Best Local Similarity 100.0%;
Matches 57; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 ATGAGGGTGTTGCTCGTTGCCCTCGCTCTCCTGGCTCTCGCTGCGAGCGCCACCAGC 57
|||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 9 ATGAGGGTGTTGCTCGTTGCCCTCGCTCTCCTGGCTCTCGCTGCGAGCGCCACCAGC 65
It would have been prima facie obvious to combine the teachings of US 2016/0340411 Al, US 20130071884 A1, and US 20030145347 A1 to design the components listed in the expression cassette as recited in Claim 47 and codon optimization to design the polynucleotide sequence in Claim 14. One having ordinary skill in the art would have a reasonable expectation of success as expression cassettes with this promoter and terminator are known in the art regarding expressing a transgene in monocots. Additionally, the maize gamma zein 27 signal peptide is known to target the protein to the seed’s endosperm and been previously used to target animal-derived protein to maize kernels1. All sequence (i.e. promoter, targeting sequence, myoglobin protein, and terminator) and techniques (i.e. codon optimization) are known in the art and considered a design choice chosen by the applicant as the criticality of these regulatory elements or sequences is not disclosed by the Applicant. One of ordinary skill in the art would have been motivated to construct an expression cassette that expresses myoglobin in corn seeds as they “a major component of human and animal diets worldwide” according to the state of the art, as evidenced by Bicar et al 2008 in paragraph 1 page 1. “Additionally, plant-based diets, such as veganism, vegetarianism, and flexitarian ism are increasing worldwide, which has generated a growing affability to plant-based diets, as well as a demand for the development of novel plant-based meat alternatives”, which could be met by Applicant’s invention2.
Citation of Relevant Prior Art
Uppal, Sheetal, et al. "Significantly enhanced heme retention ability of myoglobin engineered to mimic the third covalent linkage by nonaxial histidine to heme (vinyl) in synechocystis hemoglobin." Journal of Biological Chemistry 290.4 (2015): 1979-1993 (See IDS filed 02/07/2025) teaches the modification of myoglobin at the 107 position to a histidine for improved stability between heme and the protein (See abstract). This mutation was replicated by the Applicant in polypeptide sequence SEQ ID NO: 58.
Conclusion
No claims are allowed.
Contact Information
Any inquiry concerning this communication or earlier communications from the examiner should be directed to GEORGE W MEYER whose telephone number is (571)272-3733. The examiner can normally be reached Monday - Friday 8:00 am- 5:00 pm.
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, Bratislav Stankovic can be reached at (571)-270-0305. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. To file and manage patent submissions in Patent Center, visit: https://patentcenter.uspto.gov. Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000.
/GEORGE W MEYER/ Examiner, Art Unit 1662
/BRATISLAV STANKOVIC/ Supervisory Patent Examiner, Art Units 1661 & 1662
1 Bicar, Earl H., et al. "Transgenic maize endosperm containing a milk protein has improved amino acid balance." Transgenic research 17.1 (2008): 59-71 appears to use the same signal peptide taught by Yang, Suk-Hwan, et al. "Expression of a synthetic porcine α-lactalbumin gene in the kernels of transgenic maize." Transgenic research 11.1 (2002): 11-20 which references Prat, Salomé, et al. "Nucleic acid (cDNA) and amino acid sequences of the maize endosperm protein glutelin-2." Nucleic Acids Research 13.5 (1985): 1493-1504 which teaches an amino acid sequence of MRVLLVALALLALAASATS in Figure 4 and is 100% identical to the amino acid sequence encoded by Applicant’s SEQ ID NO:24 and described in SEQ ID NO:23.
2 Swing, Caleb J., et al. "Nutritional composition of novel plant-based meat alternatives and traditional animal-based meats." J Food Sci Nutr 7.109 (2021): 1-11.
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