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 .
Election/Restrictions
Applicant’s election without traverse of the species of SEQ ID NOs: 1, 14, 27, and 50 and “a neurodegenerative disorder” in the reply filed on 13 April 2026 is acknowledged.
Claims 73-74 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected species, there being no allowable generic or linking claim.
Claims 58-72 and 75-77 are examined herein.
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 58, 59, 60, 61, 69, and 70 are rejected under 35 U.S.C. 103 as being unpatentable over Braun et al (US20230032826 A1; Foreign Priority Date 18 Dec 2019).
Braun teaches a method for producing a recombinant viral vector from plant hairy roots, comprising inducing the formation of plant hairy roots in a plant and transforming said plants with at least one vector comprising one or more expression cassettes which comprise genes encoding the proteins required for the production of the recombinant viral vector (claim 30; instant claim 58); wherein the viral vector is adeno-associated virus (claim 35, instant claim 58); wherein the plant is an Arabidopsis (claim 31; instant claim 61).
Braun teaches that the genes encoding the proteins required for the production of the recombinant viral vector can be codon optimized to improve expression in a plant cell (¶0129; instant claim 60).
Braun teaches that plant hairy roots transformed with their recombinant AAV constructs and cultured for 20 days produce assembled AAV capsids; that said AAV capsids can be recovered (i.e. isolated) by methods which include centrifugation and said recovery results in more than 1.68 x 108 capsids/ML (see Braun “Examples”, ¶222-227, ¶235-238, ¶269-273, Table 1, and Table 2; instant claims 58, 69, and 70).
Regarding claims 58, 69 and 70 at the time of filing it would have been obvious to one of ordinary skill in the art to combine the teachings of Braun et al into a single embodiment given that Braun exemplifies a method of producing AAV particles in a Brassica rapa plant comprising transforming said plants with construct comprising sequences which encode AAV particles and all the components necessary to assemble an AAV particle in planta and given that Braun exemplifies isolating and quantifying said AAV particles, wherein in more than 1.68 x 108 capsids/ML are isolated. One would have a reasonable expectation of success given the teachings of Braun et al.
Regarding claim 59, at the time of filing it would have been obvious to one of ordinary skill in the art to further substitute one transient transformation method, i.e. agroinfiltration, for another transient transformation method, i.e. hairy root transformation. One would have been motivated to do so given that the methods are broadly interchangeable and routine in the art.
Regarding claim 60, at the time of filing it would have been obvious to one of ordinary skill in the art to further modify the method of Braun et al to include codon optimization of the nucleic acid sequences given that Braun et al teaches that the genes encoding the proteins required for the production of the recombinant viral vector can be codon optimized to improve expression in a plant cell. One would be motivated to do with a reasonable expectation of the success given the express teaching of Braun to use codon optimization and the routine nature of codon optimization for improved gene expression in plants.
Regarding claim 61, at the time of filing it would have been obvious to one of ordinary skill in the art to further modify the method of Braun et al to transform an Arabidopsis thaliana plant. One would be motivated to do with a reasonable expectation of the success given that Braun et al suggest using an Arabidopsis thaliana plant with their methods and the routine use of Arabidopsis thaliana in the art.
Claims 71 and 72 are rejected under 35 U.S.C. 103 as being unpatentable over Braun et al (US20230032826 A1; Foreign Priority Date 18 Dec 2019) as applied to claim 58 above, further in view of Blits et al (2017, Frontiers in Neuroanatomy 10: 128).
Claims 71 and 72 are drawn to the method of claim 58, further comprising administering the AAV particles to mammals, wherein the mammal comprises a neurodegenerative disorder.
The teachings of Braun et al as applied to claim 58 are set forth above.
Braun et al does not teach administering AAV particles.
Blits et al teaches that AAV mediated gene therapy has been studied for the treatment of Parkinson’s disease, a neurodegenerative disease in humans (see Table 1 and throughout).
At the time of filing, it would have been obvious to one of ordinary skill in the art to use AAV particles in a method comprising administering AAV to a mammal comprising a neurodegenerative disease. One would have been motivated to do with a reasonable expectation given that Blits et al teach AAV mediated gene therapy treatments for Parkinson’s and choosing an AAV particle produced by one method or another would constitute a substitution of equivalents.
Claims 62-63 and 65 are rejected under 35 U.S.C. 103 as being unpatentable over Braun et al (US20230032826 A1; Foreign Priority Date 18 Dec 2019) as applied to claim 58 above, further in view of Allen et al (2000, Molecular Therapy 1(1): 88-95) Maranga et al (US20210301305 A1; PCT Filed 13 June 2019).
Claims 62 is drawn to the method of claim 58, wherein the vector comprises nucleic acid sequences encoding a REP protein, a CAP protein, an AAP protein, and an AD5 E4orf6 protein.
Claims 63 and 65 are drawn to the method of claim 62 wherein the nucleic acid encoding a REP protein comprises a plant Kozak sequence that enhances translation of downstream in-frame polypeptides (claim 63) and he method of claim 62 wherein the nucleic acid encoding a CAP protein comprises a plant Kozak sequence that enhances translation of downstream in-frame polypeptides (claim 65).
The teachings of Braun et al as applied to claim 58 are set forth above.
Braun et al further teaches that the vector can comprise genes encoding VP1, VP2, VP3 (i.e. a CAP protein), AAP, Rep52 and Rep78 (i.e. a REP protein) (¶0023). Braun teaches that their method can comprise further transforming the plan with a nucleic acid sequence encoding for the adenoviral helper functions (¶0031). One of skill in the art would understand that AAV requires either a helper virus, like adenovirus, or select portions of the genome of adenovirus, like the E4 region.
Braun does not teach AD5 E4orf6.
Braun does not teach a Kozak sequence that enhances translation of downstream in-frame polypeptides.
Allen et al teaches AD5 E4orf6 is sufficient for the generation of AAV vector stocks and that this is beneficial because it eliminates the need for removing adenovirus from the AAV vector stocks (page 88, Abstract; page 89, column 1, last paragraph).
Maranga et al teaches the use of engineered UTRs which comprise weak Kozak sequences linked to a rep or cap gene, which can upregulate the expression of VP1 and VP2 relative to VP3 or downregulate VP1 and VP2 relative to VP3 or which can upregulate or downregulate the production of REP52 relative to REP78 (¶0029-0032). Maranga et al teaches that lower expression of REP78 relative to REP52 is often required for high yield production of AAV particles.
Regarding claim 62, at the time of filing, it would have been obvious to one of skill in the art to further modify the teachings of Braun et al as applied to claim 58 to include nucleic acid sequences encoding a REP protein, a CAP protein, an AAP protein, and an AD5 E4orf6 protein given the additional teachings of Braun et al and the teachings of Allen et al. One would have been motivated to include nucleic acid sequences encoding a REP protein, a CAP protein, an AAP protein, and an AD5 E4orf6 protein given that Braun teaches plans comprising VP1, VP2, VP3 (i.e. a CAP protein), AAP, Rep52 and Rep78 as well as an adenovirus helper plasmid and one of ordinary skill in the art would understand that these components are necessary for the production of functional AAV particles in a host. One would have been further motivated to replace the adenovirus helper plasmid of Braun et al with the minimal AD5 E4orf6 protein as taught by Allen et al given the explicit teachings of Allen regarding its benefits over a larger adenovirus helper plasmid.
Regarding claims 63 and 65, at the time of filing it would have been obvious to further modify the teachings of Braun et al and Allen et al to use a modified Kozak sequence given the teachings of Maranga et al. One would be motivated to do so given that the efficient assembly, production and packaging of AAV requires optimal stoichiometry of VP1:VP2:VP3 and REP78:REP52. One would have a reasonable expectation of success given the teachings of Maranga et al and given that AAV naturally uses leaky scanning to regulate the expression of multiple transcripts from the CAP gene.
Claims 64 and 66-67 are rejected under 35 U.S.C. 103 as being unpatentable over being unpatentable over Braun et al (US20230032826 A1; Foreign Priority Date 18 Dec 2019), in view of Allen et al (2000, Molecular Therapy 1(1): 88-95) and Maranga et al (US20210301305 A1; PCT Filed 13 June 2019) as applied to claims 62-63 and 65 above, and further in view of NCBI accession AF043303 (2010; .ncbi.nlm.nih.gov/nuccore/AF043303) and NCBI accession AAQ19311.1 (2007; ncbi.nlm.nih.gov/protein/AAq19311.1).
Claim 64 is drawn to the method of claim 63 wherein the REP protein is encoded by a sequence with at least 90% identity to SEQ ID NO: 1.
Claim 66 is drawn to the method of claim 65 wherein the CAP protein is encoded by a sequence with at least 90% identity to SEQ ID NO: 14.
Claim 67 is drawn to the method of claim 62 wherein the AAP protein is encoded by a sequence with at least 90% identity to SEQ ID NO: 27.
Claim 68 is drawn the method of claim 62 wherein the Ad5 E5orf6 protein comprises a sequence with at least 90% identity to SEQ ID NO: 50.
The teachings of Braun et al in view of Allen et al and Maranga et al as applied to claims as applied to claims 62-63 and 65 is set forth above.
Braun further teaches that the CAP and REP gene of AAV2 may be used. Examiner notes that the VP1, VP2, VP3, AAP, REP78, REP68, REP52 and REP40 proteins are all translated from the CAP and REP genes.
Braun does not teach SEQ ID NOs: 1, 14, 27 or 50.
NCBI accession AF043303 teaches sequences with at least 90% identity to SEQ ID NOs: 1, 14, and 27.
Alignment of instant SEQ ID NO: 1 with NCBI accession AF043303:
Query Match 99.9%; Score 1864.4; Length 4679;
Best Local Similarity 99.9%;
Matches 1865; Conservative 0; Mismatches 1; Indels 0; Gaps 0;
Qy 1 ATGCCGGGGTTTTACGAGATTGTGATTAAGGTCCCCAGCGACCTTGACGGGCATCTGCCC 60
||||||||||||||||||||||||||||||||||||||||||||||||| ||||||||||
Db 321 ATGCCGGGGTTTTACGAGATTGTGATTAAGGTCCCCAGCGACCTTGACGAGCATCTGCCC 380
Qy 61 GGCATTTCTGACAGCTTTGTGAACTGGGTGGCCGAGAAGGAATGGGAGTTGCCGCCAGAT 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 381 GGCATTTCTGACAGCTTTGTGAACTGGGTGGCCGAGAAGGAATGGGAGTTGCCGCCAGAT 440
Qy 121 TCTGACATGGATCTGAATCTGATTGAGCAGGCACCCCTGACCGTGGCCGAGAAGCTGCAG 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 441 TCTGACATGGATCTGAATCTGATTGAGCAGGCACCCCTGACCGTGGCCGAGAAGCTGCAG 500
Qy 181 CGCGACTTTCTGACGGAATGGCGCCGTGTGAGTAAGGCCCCGGAGGCCCTTTTCTTTGTG 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 501 CGCGACTTTCTGACGGAATGGCGCCGTGTGAGTAAGGCCCCGGAGGCCCTTTTCTTTGTG 560
Qy 241 CAATTTGAGAAGGGAGAGAGCTACTTCCACATGCACGTGCTCGTGGAAACCACCGGGGTG 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 561 CAATTTGAGAAGGGAGAGAGCTACTTCCACATGCACGTGCTCGTGGAAACCACCGGGGTG 620
Qy 301 AAATCCATGGTTTTGGGACGTTTCCTGAGTCAGATTCGCGAAAAACTGATTCAGAGAATT 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 621 AAATCCATGGTTTTGGGACGTTTCCTGAGTCAGATTCGCGAAAAACTGATTCAGAGAATT 680
Qy 361 TACCGCGGGATCGAGCCGACTTTGCCAAACTGGTTCGCGGTCACAAAGACCAGAAATGGC 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 681 TACCGCGGGATCGAGCCGACTTTGCCAAACTGGTTCGCGGTCACAAAGACCAGAAATGGC 740
Qy 421 GCCGGAGGCGGGAACAAGGTGGTGGATGAGTGCTACATCCCCAATTACTTGCTCCCCAAA 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 741 GCCGGAGGCGGGAACAAGGTGGTGGATGAGTGCTACATCCCCAATTACTTGCTCCCCAAA 800
Qy 481 ACCCAGCCTGAGCTCCAGTGGGCGTGGACTAATATGGAACAGTATTTAAGCGCCTGTTTG 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 801 ACCCAGCCTGAGCTCCAGTGGGCGTGGACTAATATGGAACAGTATTTAAGCGCCTGTTTG 860
Qy 541 AATCTCACGGAGCGTAAACGGTTGGTGGCGCAGCATCTGACGCACGTGTCGCAGACGCAG 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 861 AATCTCACGGAGCGTAAACGGTTGGTGGCGCAGCATCTGACGCACGTGTCGCAGACGCAG 920
Qy 601 GAGCAGAACAAAGAGAATCAGAATCCCAATTCTGATGCGCCGGTGATCAGATCAAAAACT 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 921 GAGCAGAACAAAGAGAATCAGAATCCCAATTCTGATGCGCCGGTGATCAGATCAAAAACT 980
Qy 661 TCAGCCAGGTACATGGAGCTGGTCGGGTGGCTCGTGGACAAGGGGATTACCTCGGAGAAG 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 981 TCAGCCAGGTACATGGAGCTGGTCGGGTGGCTCGTGGACAAGGGGATTACCTCGGAGAAG 1040
Qy 721 CAGTGGATCCAGGAGGACCAGGCCTCATACATCTCCTTCAATGCGGCCTCCAACTCGCGG 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1041 CAGTGGATCCAGGAGGACCAGGCCTCATACATCTCCTTCAATGCGGCCTCCAACTCGCGG 1100
Qy 781 TCCCAAATCAAGGCTGCCTTGGACAATGCGGGAAAGATTATGAGCCTGACTAAAACCGCC 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1101 TCCCAAATCAAGGCTGCCTTGGACAATGCGGGAAAGATTATGAGCCTGACTAAAACCGCC 1160
Qy 841 CCCGACTACCTGGTGGGCCAGCAGCCCGTGGAGGACATTTCCAGCAATCGGATTTATAAA 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1161 CCCGACTACCTGGTGGGCCAGCAGCCCGTGGAGGACATTTCCAGCAATCGGATTTATAAA 1220
Qy 901 ATTTTGGAACTAAACGGGTACGATCCCCAATATGCGGCTTCCGTCTTTCTGGGATGGGCC 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1221 ATTTTGGAACTAAACGGGTACGATCCCCAATATGCGGCTTCCGTCTTTCTGGGATGGGCC 1280
Qy 961 ACGAAAAAGTTCGGCAAGAGGAACACCATCTGGCTGTTTGGGCCTGCAACTACCGGGAAG 1020
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1281 ACGAAAAAGTTCGGCAAGAGGAACACCATCTGGCTGTTTGGGCCTGCAACTACCGGGAAG 1340
Qy 1021 ACCAACATCGCGGAGGCCATAGCCCACACTGTGCCCTTCTACGGGTGCGTAAACTGGACC 1080
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1341 ACCAACATCGCGGAGGCCATAGCCCACACTGTGCCCTTCTACGGGTGCGTAAACTGGACC 1400
Qy 1081 AATGAGAACTTTCCCTTCAACGACTGTGTCGACAAGATGGTGATCTGGTGGGAGGAGGGG 1140
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1401 AATGAGAACTTTCCCTTCAACGACTGTGTCGACAAGATGGTGATCTGGTGGGAGGAGGGG 1460
Qy 1141 AAGATGACCGCCAAGGTCGTGGAGTCGGCCAAAGCCATTCTCGGAGGAAGCAAGGTGCGC 1200
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1461 AAGATGACCGCCAAGGTCGTGGAGTCGGCCAAAGCCATTCTCGGAGGAAGCAAGGTGCGC 1520
Qy 1201 GTGGACCAGAAATGCAAGTCCTCGGCCCAGATAGACCCGACTCCCGTGATCGTCACCTCC 1260
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1521 GTGGACCAGAAATGCAAGTCCTCGGCCCAGATAGACCCGACTCCCGTGATCGTCACCTCC 1580
Qy 1261 AACACCAACATGTGCGCCGTGATTGACGGGAACTCAACGACCTTCGAACACCAGCAGCCG 1320
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1581 AACACCAACATGTGCGCCGTGATTGACGGGAACTCAACGACCTTCGAACACCAGCAGCCG 1640
Qy 1321 TTGCAAGACCGGATGTTCAAATTTGAACTCACCCGCCGTCTGGATCATGACTTTGGGAAG 1380
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1641 TTGCAAGACCGGATGTTCAAATTTGAACTCACCCGCCGTCTGGATCATGACTTTGGGAAG 1700
Qy 1381 GTCACCAAGCAGGAAGTCAAAGACTTTTTCCGGTGGGCAAAGGATCACGTGGTTGAGGTG 1440
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1701 GTCACCAAGCAGGAAGTCAAAGACTTTTTCCGGTGGGCAAAGGATCACGTGGTTGAGGTG 1760
Qy 1441 GAGCATGAATTCTACGTCAAAAAGGGTGGAGCCAAGAAAAGACCCGCCCCCAGTGACGCA 1500
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1761 GAGCATGAATTCTACGTCAAAAAGGGTGGAGCCAAGAAAAGACCCGCCCCCAGTGACGCA 1820
Qy 1501 GATATAAGTGAGCCCAAACGGGTGCGCGAGTCAGTTGCGCAGCCATCGACGTCAGACGCG 1560
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1821 GATATAAGTGAGCCCAAACGGGTGCGCGAGTCAGTTGCGCAGCCATCGACGTCAGACGCG 1880
Qy 1561 GAAGCTTCGATCAACTACGCAGACAGGTACCAAAACAAATGTTCTCGTCACGTGGGCATG 1620
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1881 GAAGCTTCGATCAACTACGCAGACAGGTACCAAAACAAATGTTCTCGTCACGTGGGCATG 1940
Qy 1621 AATCTGATGCTGTTTCCCTGCAGACAATGCGAGAGAATGAATCAGAATTCAAATATCTGC 1680
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1941 AATCTGATGCTGTTTCCCTGCAGACAATGCGAGAGAATGAATCAGAATTCAAATATCTGC 2000
Qy 1681 TTCACTCACGGACAGAAAGACTGTTTAGAGTGCTTTCCCGTGTCAGAATCTCAACCCGTT 1740
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2001 TTCACTCACGGACAGAAAGACTGTTTAGAGTGCTTTCCCGTGTCAGAATCTCAACCCGTT 2060
Qy 1741 TCTGTCGTCAAAAAGGCGTATCAGAAACTGTGCTACATTCATCATATCATGGGAAAGGTG 1800
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2061 TCTGTCGTCAAAAAGGCGTATCAGAAACTGTGCTACATTCATCATATCATGGGAAAGGTG 2120
Qy 1801 CCAGACGCTTGCACTGCCTGCGATCTGGTCAATGTGGATTTGGATGACTGCATCTTTGAA 1860
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2121 CCAGACGCTTGCACTGCCTGCGATCTGGTCAATGTGGATTTGGATGACTGCATCTTTGAA 2180
Qy 1861 CAATAA 1866
||||||
Db 2181 CAATAA 2186
Alignment of instant SEQ ID NO: 14 with NCBI accession AF043303:
Query Match 100.0%; Score 2208; Length 4679;
Best Local Similarity 100.0%;
Matches 2208; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 ATGGCTGCCGATGGTTATCTTCCAGATTGGCTCGAGGACACTCTCTCTGAAGGAATAAGA 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2203 ATGGCTGCCGATGGTTATCTTCCAGATTGGCTCGAGGACACTCTCTCTGAAGGAATAAGA 2262
Qy 61 CAGTGGTGGAAGCTCAAACCTGGCCCACCACCACCAAAGCCCGCAGAGCGGCATAAGGAC 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2263 CAGTGGTGGAAGCTCAAACCTGGCCCACCACCACCAAAGCCCGCAGAGCGGCATAAGGAC 2322
Qy 121 GACAGCAGGGGTCTTGTGCTTCCTGGGTACAAGTACCTCGGACCCTTCAACGGACTCGAC 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2323 GACAGCAGGGGTCTTGTGCTTCCTGGGTACAAGTACCTCGGACCCTTCAACGGACTCGAC 2382
Qy 181 AAGGGAGAGCCGGTCAACGAGGCAGACGCCGCGGCCCTCGAGCACGACAAAGCCTACGAC 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2383 AAGGGAGAGCCGGTCAACGAGGCAGACGCCGCGGCCCTCGAGCACGACAAAGCCTACGAC 2442
Qy 241 CGGCAGCTCGACAGCGGAGACAACCCGTACCTCAAGTACAACCACGCCGACGCGGAGTTT 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2443 CGGCAGCTCGACAGCGGAGACAACCCGTACCTCAAGTACAACCACGCCGACGCGGAGTTT 2502
Qy 301 CAGGAGCGCCTTAAAGAAGATACGTCTTTTGGGGGCAACCTCGGACGAGCAGTCTTCCAG 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2503 CAGGAGCGCCTTAAAGAAGATACGTCTTTTGGGGGCAACCTCGGACGAGCAGTCTTCCAG 2562
Qy 361 GCGAAAAAGAGGGTTCTTGAACCTCTGGGCCTGGTTGAGGAACCTGTTAAGACGGCTCCG 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2563 GCGAAAAAGAGGGTTCTTGAACCTCTGGGCCTGGTTGAGGAACCTGTTAAGACGGCTCCG 2622
Qy 421 GGAAAAAAGAGGCCGGTAGAGCACTCTCCTGTGGAGCCAGACTCCTCCTCGGGAACCGGA 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2623 GGAAAAAAGAGGCCGGTAGAGCACTCTCCTGTGGAGCCAGACTCCTCCTCGGGAACCGGA 2682
Qy 481 AAGGCGGGCCAGCAGCCTGCAAGAAAAAGATTGAATTTTGGTCAGACTGGAGACGCAGAC 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2683 AAGGCGGGCCAGCAGCCTGCAAGAAAAAGATTGAATTTTGGTCAGACTGGAGACGCAGAC 2742
Qy 541 TCAGTACCTGACCCCCAGCCTCTCGGACAGCCACCAGCAGCCCCCTCTGGTCTGGGAACT 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2743 TCAGTACCTGACCCCCAGCCTCTCGGACAGCCACCAGCAGCCCCCTCTGGTCTGGGAACT 2802
Qy 601 AATACGATGGCTACAGGCAGTGGCGCACCAATGGCAGACAATAACGAGGGCGCCGACGGA 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2803 AATACGATGGCTACAGGCAGTGGCGCACCAATGGCAGACAATAACGAGGGCGCCGACGGA 2862
Qy 661 GTGGGTAATTCCTCGGGAAATTGGCATTGCGATTCCACATGGATGGGCGACAGAGTCATC 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2863 GTGGGTAATTCCTCGGGAAATTGGCATTGCGATTCCACATGGATGGGCGACAGAGTCATC 2922
Qy 721 ACCACCAGCACCCGAACCTGGGCCCTGCCCACCTACAACAACCACCTCTACAAACAAATT 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2923 ACCACCAGCACCCGAACCTGGGCCCTGCCCACCTACAACAACCACCTCTACAAACAAATT 2982
Qy 781 TCCAGCCAATCAGGAGCCTCGAACGACAATCACTACTTTGGCTACAGCACCCCTTGGGGG 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2983 TCCAGCCAATCAGGAGCCTCGAACGACAATCACTACTTTGGCTACAGCACCCCTTGGGGG 3042
Qy 841 TATTTTGACTTCAACAGATTCCACTGCCACTTTTCACCACGTGACTGGCAAAGACTCATC 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3043 TATTTTGACTTCAACAGATTCCACTGCCACTTTTCACCACGTGACTGGCAAAGACTCATC 3102
Qy 901 AACAACAACTGGGGATTCCGACCCAAGAGACTCAACTTCAAGCTCTTTAACATTCAAGTC 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3103 AACAACAACTGGGGATTCCGACCCAAGAGACTCAACTTCAAGCTCTTTAACATTCAAGTC 3162
Qy 961 AAAGAGGTCACGCAGAATGACGGTACGACGACGATTGCCAATAACCTTACCAGCACGGTT 1020
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3163 AAAGAGGTCACGCAGAATGACGGTACGACGACGATTGCCAATAACCTTACCAGCACGGTT 3222
Qy 1021 CAGGTGTTTACTGACTCGGAGTACCAGCTCCCGTACGTCCTCGGCTCGGCGCATCAAGGA 1080
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3223 CAGGTGTTTACTGACTCGGAGTACCAGCTCCCGTACGTCCTCGGCTCGGCGCATCAAGGA 3282
Qy 1081 TGCCTCCCGCCGTTCCCAGCAGACGTCTTCATGGTGCCACAGTATGGATACCTCACCCTG 1140
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3283 TGCCTCCCGCCGTTCCCAGCAGACGTCTTCATGGTGCCACAGTATGGATACCTCACCCTG 3342
Qy 1141 AACAACGGGAGTCAGGCAGTAGGACGCTCTTCATTTTACTGCCTGGAGTACTTTCCTTCT 1200
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3343 AACAACGGGAGTCAGGCAGTAGGACGCTCTTCATTTTACTGCCTGGAGTACTTTCCTTCT 3402
Qy 1201 CAGATGCTGCGTACCGGAAACAACTTTACCTTCAGCTACACTTTTGAGGACGTTCCTTTC 1260
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3403 CAGATGCTGCGTACCGGAAACAACTTTACCTTCAGCTACACTTTTGAGGACGTTCCTTTC 3462
Qy 1261 CACAGCAGCTACGCTCACAGCCAGAGTCTGGACCGTCTCATGAATCCTCTCATCGACCAG 1320
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3463 CACAGCAGCTACGCTCACAGCCAGAGTCTGGACCGTCTCATGAATCCTCTCATCGACCAG 3522
Qy 1321 TACCTGTATTACTTGAGCAGAACAAACACTCCAAGTGGAACCACCACGCAGTCAAGGCTT 1380
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3523 TACCTGTATTACTTGAGCAGAACAAACACTCCAAGTGGAACCACCACGCAGTCAAGGCTT 3582
Qy 1381 CAGTTTTCTCAGGCCGGAGCGAGTGACATTCGGGACCAGTCTAGGAACTGGCTTCCTGGA 1440
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3583 CAGTTTTCTCAGGCCGGAGCGAGTGACATTCGGGACCAGTCTAGGAACTGGCTTCCTGGA 3642
Qy 1441 CCCTGTTACCGCCAGCAGCGAGTATCAAAGACATCTGCGGATAACAACAACAGTGAATAC 1500
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3643 CCCTGTTACCGCCAGCAGCGAGTATCAAAGACATCTGCGGATAACAACAACAGTGAATAC 3702
Qy 1501 TCGTGGACTGGAGCTACCAAGTACCACCTCAATGGCAGAGACTCTCTGGTGAATCCGGGC 1560
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3703 TCGTGGACTGGAGCTACCAAGTACCACCTCAATGGCAGAGACTCTCTGGTGAATCCGGGC 3762
Qy 1561 CCGGCCATGGCAAGCCACAAGGACGATGAAGAAAAGTTTTTTCCTCAGAGCGGGGTTCTC 1620
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3763 CCGGCCATGGCAAGCCACAAGGACGATGAAGAAAAGTTTTTTCCTCAGAGCGGGGTTCTC 3822
Qy 1621 ATCTTTGGGAAGCAAGGCTCAGAGAAAACAAATGTGGACATTGAAAAGGTCATGATTACA 1680
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3823 ATCTTTGGGAAGCAAGGCTCAGAGAAAACAAATGTGGACATTGAAAAGGTCATGATTACA 3882
Qy 1681 GACGAAGAGGAAATCAGGACAACCAATCCCGTGGCTACGGAGCAGTATGGTTCTGTATCT 1740
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3883 GACGAAGAGGAAATCAGGACAACCAATCCCGTGGCTACGGAGCAGTATGGTTCTGTATCT 3942
Qy 1741 ACCAACCTCCAGAGAGGCAACAGACAAGCAGCTACCGCAGATGTCAACACACAAGGCGTT 1800
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3943 ACCAACCTCCAGAGAGGCAACAGACAAGCAGCTACCGCAGATGTCAACACACAAGGCGTT 4002
Qy 1801 CTTCCAGGCATGGTCTGGCAGGACAGAGATGTGTACCTTCAGGGGCCCATCTGGGCAAAG 1860
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4003 CTTCCAGGCATGGTCTGGCAGGACAGAGATGTGTACCTTCAGGGGCCCATCTGGGCAAAG 4062
Qy 1861 ATTCCACACACGGACGGACATTTTCACCCCTCTCCCCTCATGGGTGGATTCGGACTTAAA 1920
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4063 ATTCCACACACGGACGGACATTTTCACCCCTCTCCCCTCATGGGTGGATTCGGACTTAAA 4122
Qy 1921 CACCCTCCTCCACAGATTCTCATCAAGAACACCCCGGTACCTGCGAATCCTTCGACCACC 1980
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4123 CACCCTCCTCCACAGATTCTCATCAAGAACACCCCGGTACCTGCGAATCCTTCGACCACC 4182
Qy 1981 TTCAGTGCGGCAAAGTTTGCTTCCTTCATCACACAGTACTCCACGGGACAGGTCAGCGTG 2040
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4183 TTCAGTGCGGCAAAGTTTGCTTCCTTCATCACACAGTACTCCACGGGACAGGTCAGCGTG 4242
Qy 2041 GAGATCGAGTGGGAGCTGCAGAAGGAAAACAGCAAACGCTGGAATCCCGAAATTCAGTAC 2100
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4243 GAGATCGAGTGGGAGCTGCAGAAGGAAAACAGCAAACGCTGGAATCCCGAAATTCAGTAC 4302
Qy 2101 ACTTCCAACTACAACAAGTCTGTTAATGTGGACTTTACTGTGGACACTAATGGCGTGTAT 2160
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4303 ACTTCCAACTACAACAAGTCTGTTAATGTGGACTTTACTGTGGACACTAATGGCGTGTAT 4362
Qy 2161 TCAGAGCCTCGCCCCATTGGCACCAGATACCTGACTCGTAATCTGTAA 2208
||||||||||||||||||||||||||||||||||||||||||||||||
Db 4363 TCAGAGCCTCGCCCCATTGGCACCAGATACCTGACTCGTAATCTGTAA 4410
Alignment of instant SEQ ID NO: 27 with NCBI accession AF043303:
Query Match 99.5%; Score 615; Length 2208;
Best Local Similarity 100.0%;
Matches 615; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 4 CTGGAGACGCAGACTCAGTACCTGACCCCCAGCCTCTCGGACAGCCACCAGCAGCCCCCT 63
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 527 CTGGAGACGCAGACTCAGTACCTGACCCCCAGCCTCTCGGACAGCCACCAGCAGCCCCCT 586
Qy 64 CTGGTCTGGGAACTAATACGATGGCTACAGGCAGTGGCGCACCAATGGCAGACAATAACG 123
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 587 CTGGTCTGGGAACTAATACGATGGCTACAGGCAGTGGCGCACCAATGGCAGACAATAACG 646
Qy 124 AGGGCGCCGACGGAGTGGGTAATTCCTCGGGAAATTGGCATTGCGATTCCACATGGATGG 183
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 647 AGGGCGCCGACGGAGTGGGTAATTCCTCGGGAAATTGGCATTGCGATTCCACATGGATGG 706
Qy 184 GCGACAGAGTCATCACCACCAGCACCCGAACCTGGGCCCTGCCCACCTACAACAACCACC 243
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 707 GCGACAGAGTCATCACCACCAGCACCCGAACCTGGGCCCTGCCCACCTACAACAACCACC 766
Qy 244 TCTACAAACAAATTTCCAGCCAATCAGGAGCCTCGAACGACAATCACTACTTTGGCTACA 303
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 767 TCTACAAACAAATTTCCAGCCAATCAGGAGCCTCGAACGACAATCACTACTTTGGCTACA 826
Qy 304 GCACCCCTTGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTTTCACCACGTGACT 363
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 827 GCACCCCTTGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTTTCACCACGTGACT 886
Qy 364 GGCAAAGACTCATCAACAACAACTGGGGATTCCGACCCAAGAGACTCAACTTCAAGCTCT 423
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 887 GGCAAAGACTCATCAACAACAACTGGGGATTCCGACCCAAGAGACTCAACTTCAAGCTCT 946
Qy 424 TTAACATTCAAGTCAAAGAGGTCACGCAGAATGACGGTACGACGACGATTGCCAATAACC 483
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 947 TTAACATTCAAGTCAAAGAGGTCACGCAGAATGACGGTACGACGACGATTGCCAATAACC 1006
Qy 484 TTACCAGCACGGTTCAGGTGTTTACTGACTCGGAGTACCAGCTCCCGTACGTCCTCGGCT 543
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1007 TTACCAGCACGGTTCAGGTGTTTACTGACTCGGAGTACCAGCTCCCGTACGTCCTCGGCT 1066
Qy 544 CGGCGCATCAAGGATGCCTCCCGCCGTTCCCAGCAGACGTCTTCATGGTGCCACAGTATG 603
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1067 CGGCGCATCAAGGATGCCTCCCGCCGTTCCCAGCAGACGTCTTCATGGTGCCACAGTATG 1126
Qy 604 GATACCTCACCCTGA 618
|||||||||||||||
Db 1127 GATACCTCACCCTGA 1141
NCBI accession AAQ19311.1 teaches a sequence with 90% identity to SEQ ID NO: 50.
Alignment of instant SEQ ID NO: 50 with NCBI accession AAQ19311.1:
Query Match 100.0%; Score 799; Length 150;
Best Local Similarity 100.0%;
Matches 150; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MTTSGVPFGMTLRPTRSRLSRRTPYSRDRLPPFETETRATILEDHPLLPECNTLTMHNAW 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MTTSGVPFGMTLRPTRSRLSRRTPYSRDRLPPFETETRATILEDHPLLPECNTLTMHNAW 60
Qy 61 TSPSPPVKQPQVGQQPVAQQLDSDMNLSELPGEFINITDERLARQETVWNITPKNMSVTH 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 TSPSPPVKQPQVGQQPVAQQLDSDMNLSELPGEFINITDERLARQETVWNITPKNMSVTH 120
Qy 121 DMMLFKASRGERTVYSVCWEGGGRLNTRVL 150
||||||||||||||||||||||||||||||
Db 121 DMMLFKASRGERTVYSVCWEGGGRLNTRVL 150
At the time of filing, it would have been obvious to one of ordinary skill in the art to further modify the combined teachings of Braun et al, Allen et al, and Maranga et al to select the wild type AAV2 sequences disclosed by NCBI accession AF043303 and the E4orf6 sequence disclosed by AAQ19311.1 given the explicit suggestions of Braun et al that AAV2 sequences can be used to produce recombinant AAV in plant cells and given the teachings of Allen et al to use E4orf6.
Claims 75-77 are rejected under 35 U.S.C. 103 as being unpatentable over Braun et al (US20230032826 A1; Foreign Priority Date 18 Dec 2019) in view of Allen et al (2000, Molecular Therapy 1(1): 88-95), NCBI accession AF043303 (2010; .ncbi.nlm.nih.gov/nuccore/AF043303) and NCBI accession AAQ19311.1 (2007; ncbi.nlm.nih.gov/protein/AAq19311.1).
Braun teaches a method for producing a recombinant viral vector from plant hairy roots, comprising inducing the formation of plant hairy roots in a plant and transforming said plants with at least one vector comprising one or more expression cassettes which comprise genes encoding the proteins required for the production of the recombinant viral vector (claim 30); wherein the viral vector is adeno-associated virus (claim 35); wherein the plant is an Arabidopsis (claim 31).
Braun teaches that the genes encoding the proteins required for the production of the recombinant viral vector can be codon optimized to improve expression in a plant cell (¶0129).
Braun et al further teaches that the vector can comprise genes encoding VP1, VP2, VP3 (i.e. a CAP protein), AAP, Rep52 and Rep78 (i.e. a REP protein) (¶0023). Braun teaches that their method can comprise further transforming the plan with a nucleic acid sequence encoding for the adenoviral helper functions (¶0031). One of skill in the art would understand that AAV requires either a helper virus, like adenovirus, or select portions of the genome of adenovirus, like the E4 region.
Braun further teaches that the CAP and REP gene of AAV2 may be used. Examiner notes that the VP1, VP2, VP3, AAP, REP78, REP68, REP52 and REP40 proteins are all translated from the CAP and REP genes.
Braun teaches that plant hairy roots transformed with their recombinant AAV constructs and cultured for 20 days produce assembled AAV capsids; that said AAV capsids can be recovered (i.e. isolated) by methods which include centrifugation and said recovery results in more than 1.68 x 108 capsids/ML (see Braun “Examples”, ¶222-227, ¶235-238, ¶269-273, Table 1, and Table 2; instant claims 58, 69, and 70).
Braun does not teach AD5 E4orf6.
Braun does not teach SEQ ID NOs: 1, 14, 27 or 50.
Allen et al teaches AD5 E4orf6 is sufficient for the generation of AAV vector stocks and that this is beneficial because it eliminates the need for removing adenovirus from the AAV vector stocks (page 88, Abstract; page 89, column 1, last paragraph).
NCBI accession AF043303 teaches sequences with at least 90% identity to SEQ ID NOs: 1, 14, and 27.
Alignment of instant SEQ ID NO: 1 with NCBI accession AF043303:
Query Match 99.9%; Score 1864.4; Length 4679;
Best Local Similarity 99.9%;
Matches 1865; Conservative 0; Mismatches 1; Indels 0; Gaps 0;
Qy 1 ATGCCGGGGTTTTACGAGATTGTGATTAAGGTCCCCAGCGACCTTGACGGGCATCTGCCC 60
||||||||||||||||||||||||||||||||||||||||||||||||| ||||||||||
Db 321 ATGCCGGGGTTTTACGAGATTGTGATTAAGGTCCCCAGCGACCTTGACGAGCATCTGCCC 380
Qy 61 GGCATTTCTGACAGCTTTGTGAACTGGGTGGCCGAGAAGGAATGGGAGTTGCCGCCAGAT 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 381 GGCATTTCTGACAGCTTTGTGAACTGGGTGGCCGAGAAGGAATGGGAGTTGCCGCCAGAT 440
Qy 121 TCTGACATGGATCTGAATCTGATTGAGCAGGCACCCCTGACCGTGGCCGAGAAGCTGCAG 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 441 TCTGACATGGATCTGAATCTGATTGAGCAGGCACCCCTGACCGTGGCCGAGAAGCTGCAG 500
Qy 181 CGCGACTTTCTGACGGAATGGCGCCGTGTGAGTAAGGCCCCGGAGGCCCTTTTCTTTGTG 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 501 CGCGACTTTCTGACGGAATGGCGCCGTGTGAGTAAGGCCCCGGAGGCCCTTTTCTTTGTG 560
Qy 241 CAATTTGAGAAGGGAGAGAGCTACTTCCACATGCACGTGCTCGTGGAAACCACCGGGGTG 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 561 CAATTTGAGAAGGGAGAGAGCTACTTCCACATGCACGTGCTCGTGGAAACCACCGGGGTG 620
Qy 301 AAATCCATGGTTTTGGGACGTTTCCTGAGTCAGATTCGCGAAAAACTGATTCAGAGAATT 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 621 AAATCCATGGTTTTGGGACGTTTCCTGAGTCAGATTCGCGAAAAACTGATTCAGAGAATT 680
Qy 361 TACCGCGGGATCGAGCCGACTTTGCCAAACTGGTTCGCGGTCACAAAGACCAGAAATGGC 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 681 TACCGCGGGATCGAGCCGACTTTGCCAAACTGGTTCGCGGTCACAAAGACCAGAAATGGC 740
Qy 421 GCCGGAGGCGGGAACAAGGTGGTGGATGAGTGCTACATCCCCAATTACTTGCTCCCCAAA 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 741 GCCGGAGGCGGGAACAAGGTGGTGGATGAGTGCTACATCCCCAATTACTTGCTCCCCAAA 800
Qy 481 ACCCAGCCTGAGCTCCAGTGGGCGTGGACTAATATGGAACAGTATTTAAGCGCCTGTTTG 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 801 ACCCAGCCTGAGCTCCAGTGGGCGTGGACTAATATGGAACAGTATTTAAGCGCCTGTTTG 860
Qy 541 AATCTCACGGAGCGTAAACGGTTGGTGGCGCAGCATCTGACGCACGTGTCGCAGACGCAG 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 861 AATCTCACGGAGCGTAAACGGTTGGTGGCGCAGCATCTGACGCACGTGTCGCAGACGCAG 920
Qy 601 GAGCAGAACAAAGAGAATCAGAATCCCAATTCTGATGCGCCGGTGATCAGATCAAAAACT 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 921 GAGCAGAACAAAGAGAATCAGAATCCCAATTCTGATGCGCCGGTGATCAGATCAAAAACT 980
Qy 661 TCAGCCAGGTACATGGAGCTGGTCGGGTGGCTCGTGGACAAGGGGATTACCTCGGAGAAG 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 981 TCAGCCAGGTACATGGAGCTGGTCGGGTGGCTCGTGGACAAGGGGATTACCTCGGAGAAG 1040
Qy 721 CAGTGGATCCAGGAGGACCAGGCCTCATACATCTCCTTCAATGCGGCCTCCAACTCGCGG 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1041 CAGTGGATCCAGGAGGACCAGGCCTCATACATCTCCTTCAATGCGGCCTCCAACTCGCGG 1100
Qy 781 TCCCAAATCAAGGCTGCCTTGGACAATGCGGGAAAGATTATGAGCCTGACTAAAACCGCC 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1101 TCCCAAATCAAGGCTGCCTTGGACAATGCGGGAAAGATTATGAGCCTGACTAAAACCGCC 1160
Qy 841 CCCGACTACCTGGTGGGCCAGCAGCCCGTGGAGGACATTTCCAGCAATCGGATTTATAAA 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1161 CCCGACTACCTGGTGGGCCAGCAGCCCGTGGAGGACATTTCCAGCAATCGGATTTATAAA 1220
Qy 901 ATTTTGGAACTAAACGGGTACGATCCCCAATATGCGGCTTCCGTCTTTCTGGGATGGGCC 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1221 ATTTTGGAACTAAACGGGTACGATCCCCAATATGCGGCTTCCGTCTTTCTGGGATGGGCC 1280
Qy 961 ACGAAAAAGTTCGGCAAGAGGAACACCATCTGGCTGTTTGGGCCTGCAACTACCGGGAAG 1020
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1281 ACGAAAAAGTTCGGCAAGAGGAACACCATCTGGCTGTTTGGGCCTGCAACTACCGGGAAG 1340
Qy 1021 ACCAACATCGCGGAGGCCATAGCCCACACTGTGCCCTTCTACGGGTGCGTAAACTGGACC 1080
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1341 ACCAACATCGCGGAGGCCATAGCCCACACTGTGCCCTTCTACGGGTGCGTAAACTGGACC 1400
Qy 1081 AATGAGAACTTTCCCTTCAACGACTGTGTCGACAAGATGGTGATCTGGTGGGAGGAGGGG 1140
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1401 AATGAGAACTTTCCCTTCAACGACTGTGTCGACAAGATGGTGATCTGGTGGGAGGAGGGG 1460
Qy 1141 AAGATGACCGCCAAGGTCGTGGAGTCGGCCAAAGCCATTCTCGGAGGAAGCAAGGTGCGC 1200
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1461 AAGATGACCGCCAAGGTCGTGGAGTCGGCCAAAGCCATTCTCGGAGGAAGCAAGGTGCGC 1520
Qy 1201 GTGGACCAGAAATGCAAGTCCTCGGCCCAGATAGACCCGACTCCCGTGATCGTCACCTCC 1260
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1521 GTGGACCAGAAATGCAAGTCCTCGGCCCAGATAGACCCGACTCCCGTGATCGTCACCTCC 1580
Qy 1261 AACACCAACATGTGCGCCGTGATTGACGGGAACTCAACGACCTTCGAACACCAGCAGCCG 1320
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1581 AACACCAACATGTGCGCCGTGATTGACGGGAACTCAACGACCTTCGAACACCAGCAGCCG 1640
Qy 1321 TTGCAAGACCGGATGTTCAAATTTGAACTCACCCGCCGTCTGGATCATGACTTTGGGAAG 1380
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1641 TTGCAAGACCGGATGTTCAAATTTGAACTCACCCGCCGTCTGGATCATGACTTTGGGAAG 1700
Qy 1381 GTCACCAAGCAGGAAGTCAAAGACTTTTTCCGGTGGGCAAAGGATCACGTGGTTGAGGTG 1440
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1701 GTCACCAAGCAGGAAGTCAAAGACTTTTTCCGGTGGGCAAAGGATCACGTGGTTGAGGTG 1760
Qy 1441 GAGCATGAATTCTACGTCAAAAAGGGTGGAGCCAAGAAAAGACCCGCCCCCAGTGACGCA 1500
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1761 GAGCATGAATTCTACGTCAAAAAGGGTGGAGCCAAGAAAAGACCCGCCCCCAGTGACGCA 1820
Qy 1501 GATATAAGTGAGCCCAAACGGGTGCGCGAGTCAGTTGCGCAGCCATCGACGTCAGACGCG 1560
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1821 GATATAAGTGAGCCCAAACGGGTGCGCGAGTCAGTTGCGCAGCCATCGACGTCAGACGCG 1880
Qy 1561 GAAGCTTCGATCAACTACGCAGACAGGTACCAAAACAAATGTTCTCGTCACGTGGGCATG 1620
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1881 GAAGCTTCGATCAACTACGCAGACAGGTACCAAAACAAATGTTCTCGTCACGTGGGCATG 1940
Qy 1621 AATCTGATGCTGTTTCCCTGCAGACAATGCGAGAGAATGAATCAGAATTCAAATATCTGC 1680
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1941 AATCTGATGCTGTTTCCCTGCAGACAATGCGAGAGAATGAATCAGAATTCAAATATCTGC 2000
Qy 1681 TTCACTCACGGACAGAAAGACTGTTTAGAGTGCTTTCCCGTGTCAGAATCTCAACCCGTT 1740
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2001 TTCACTCACGGACAGAAAGACTGTTTAGAGTGCTTTCCCGTGTCAGAATCTCAACCCGTT 2060
Qy 1741 TCTGTCGTCAAAAAGGCGTATCAGAAACTGTGCTACATTCATCATATCATGGGAAAGGTG 1800
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2061 TCTGTCGTCAAAAAGGCGTATCAGAAACTGTGCTACATTCATCATATCATGGGAAAGGTG 2120
Qy 1801 CCAGACGCTTGCACTGCCTGCGATCTGGTCAATGTGGATTTGGATGACTGCATCTTTGAA 1860
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2121 CCAGACGCTTGCACTGCCTGCGATCTGGTCAATGTGGATTTGGATGACTGCATCTTTGAA 2180
Qy 1861 CAATAA 1866
||||||
Db 2181 CAATAA 2186
Alignment of instant SEQ ID NO: 14 with NCBI accession AF043303:
Query Match 100.0%; Score 2208; Length 4679;
Best Local Similarity 100.0%;
Matches 2208; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 ATGGCTGCCGATGGTTATCTTCCAGATTGGCTCGAGGACACTCTCTCTGAAGGAATAAGA 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2203 ATGGCTGCCGATGGTTATCTTCCAGATTGGCTCGAGGACACTCTCTCTGAAGGAATAAGA 2262
Qy 61 CAGTGGTGGAAGCTCAAACCTGGCCCACCACCACCAAAGCCCGCAGAGCGGCATAAGGAC 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2263 CAGTGGTGGAAGCTCAAACCTGGCCCACCACCACCAAAGCCCGCAGAGCGGCATAAGGAC 2322
Qy 121 GACAGCAGGGGTCTTGTGCTTCCTGGGTACAAGTACCTCGGACCCTTCAACGGACTCGAC 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2323 GACAGCAGGGGTCTTGTGCTTCCTGGGTACAAGTACCTCGGACCCTTCAACGGACTCGAC 2382
Qy 181 AAGGGAGAGCCGGTCAACGAGGCAGACGCCGCGGCCCTCGAGCACGACAAAGCCTACGAC 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2383 AAGGGAGAGCCGGTCAACGAGGCAGACGCCGCGGCCCTCGAGCACGACAAAGCCTACGAC 2442
Qy 241 CGGCAGCTCGACAGCGGAGACAACCCGTACCTCAAGTACAACCACGCCGACGCGGAGTTT 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2443 CGGCAGCTCGACAGCGGAGACAACCCGTACCTCAAGTACAACCACGCCGACGCGGAGTTT 2502
Qy 301 CAGGAGCGCCTTAAAGAAGATACGTCTTTTGGGGGCAACCTCGGACGAGCAGTCTTCCAG 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2503 CAGGAGCGCCTTAAAGAAGATACGTCTTTTGGGGGCAACCTCGGACGAGCAGTCTTCCAG 2562
Qy 361 GCGAAAAAGAGGGTTCTTGAACCTCTGGGCCTGGTTGAGGAACCTGTTAAGACGGCTCCG 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2563 GCGAAAAAGAGGGTTCTTGAACCTCTGGGCCTGGTTGAGGAACCTGTTAAGACGGCTCCG 2622
Qy 421 GGAAAAAAGAGGCCGGTAGAGCACTCTCCTGTGGAGCCAGACTCCTCCTCGGGAACCGGA 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2623 GGAAAAAAGAGGCCGGTAGAGCACTCTCCTGTGGAGCCAGACTCCTCCTCGGGAACCGGA 2682
Qy 481 AAGGCGGGCCAGCAGCCTGCAAGAAAAAGATTGAATTTTGGTCAGACTGGAGACGCAGAC 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2683 AAGGCGGGCCAGCAGCCTGCAAGAAAAAGATTGAATTTTGGTCAGACTGGAGACGCAGAC 2742
Qy 541 TCAGTACCTGACCCCCAGCCTCTCGGACAGCCACCAGCAGCCCCCTCTGGTCTGGGAACT 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2743 TCAGTACCTGACCCCCAGCCTCTCGGACAGCCACCAGCAGCCCCCTCTGGTCTGGGAACT 2802
Qy 601 AATACGATGGCTACAGGCAGTGGCGCACCAATGGCAGACAATAACGAGGGCGCCGACGGA 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2803 AATACGATGGCTACAGGCAGTGGCGCACCAATGGCAGACAATAACGAGGGCGCCGACGGA 2862
Qy 661 GTGGGTAATTCCTCGGGAAATTGGCATTGCGATTCCACATGGATGGGCGACAGAGTCATC 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2863 GTGGGTAATTCCTCGGGAAATTGGCATTGCGATTCCACATGGATGGGCGACAGAGTCATC 2922
Qy 721 ACCACCAGCACCCGAACCTGGGCCCTGCCCACCTACAACAACCACCTCTACAAACAAATT 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2923 ACCACCAGCACCCGAACCTGGGCCCTGCCCACCTACAACAACCACCTCTACAAACAAATT 2982
Qy 781 TCCAGCCAATCAGGAGCCTCGAACGACAATCACTACTTTGGCTACAGCACCCCTTGGGGG 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2983 TCCAGCCAATCAGGAGCCTCGAACGACAATCACTACTTTGGCTACAGCACCCCTTGGGGG 3042
Qy 841 TATTTTGACTTCAACAGATTCCACTGCCACTTTTCACCACGTGACTGGCAAAGACTCATC 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3043 TATTTTGACTTCAACAGATTCCACTGCCACTTTTCACCACGTGACTGGCAAAGACTCATC 3102
Qy 901 AACAACAACTGGGGATTCCGACCCAAGAGACTCAACTTCAAGCTCTTTAACATTCAAGTC 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3103 AACAACAACTGGGGATTCCGACCCAAGAGACTCAACTTCAAGCTCTTTAACATTCAAGTC 3162
Qy 961 AAAGAGGTCACGCAGAATGACGGTACGACGACGATTGCCAATAACCTTACCAGCACGGTT 1020
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3163 AAAGAGGTCACGCAGAATGACGGTACGACGACGATTGCCAATAACCTTACCAGCACGGTT 3222
Qy 1021 CAGGTGTTTACTGACTCGGAGTACCAGCTCCCGTACGTCCTCGGCTCGGCGCATCAAGGA 1080
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3223 CAGGTGTTTACTGACTCGGAGTACCAGCTCCCGTACGTCCTCGGCTCGGCGCATCAAGGA 3282
Qy 1081 TGCCTCCCGCCGTTCCCAGCAGACGTCTTCATGGTGCCACAGTATGGATACCTCACCCTG 1140
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3283 TGCCTCCCGCCGTTCCCAGCAGACGTCTTCATGGTGCCACAGTATGGATACCTCACCCTG 3342
Qy 1141 AACAACGGGAGTCAGGCAGTAGGACGCTCTTCATTTTACTGCCTGGAGTACTTTCCTTCT 1200
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3343 AACAACGGGAGTCAGGCAGTAGGACGCTCTTCATTTTACTGCCTGGAGTACTTTCCTTCT 3402
Qy 1201 CAGATGCTGCGTACCGGAAACAACTTTACCTTCAGCTACACTTTTGAGGACGTTCCTTTC 1260
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3403 CAGATGCTGCGTACCGGAAACAACTTTACCTTCAGCTACACTTTTGAGGACGTTCCTTTC 3462
Qy 1261 CACAGCAGCTACGCTCACAGCCAGAGTCTGGACCGTCTCATGAATCCTCTCATCGACCAG 1320
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3463 CACAGCAGCTACGCTCACAGCCAGAGTCTGGACCGTCTCATGAATCCTCTCATCGACCAG 3522
Qy 1321 TACCTGTATTACTTGAGCAGAACAAACACTCCAAGTGGAACCACCACGCAGTCAAGGCTT 1380
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3523 TACCTGTATTACTTGAGCAGAACAAACACTCCAAGTGGAACCACCACGCAGTCAAGGCTT 3582
Qy 1381 CAGTTTTCTCAGGCCGGAGCGAGTGACATTCGGGACCAGTCTAGGAACTGGCTTCCTGGA 1440
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3583 CAGTTTTCTCAGGCCGGAGCGAGTGACATTCGGGACCAGTCTAGGAACTGGCTTCCTGGA 3642
Qy 1441 CCCTGTTACCGCCAGCAGCGAGTATCAAAGACATCTGCGGATAACAACAACAGTGAATAC 1500
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3643 CCCTGTTACCGCCAGCAGCGAGTATCAAAGACATCTGCGGATAACAACAACAGTGAATAC 3702
Qy 1501 TCGTGGACTGGAGCTACCAAGTACCACCTCAATGGCAGAGACTCTCTGGTGAATCCGGGC 1560
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3703 TCGTGGACTGGAGCTACCAAGTACCACCTCAATGGCAGAGACTCTCTGGTGAATCCGGGC 3762
Qy 1561 CCGGCCATGGCAAGCCACAAGGACGATGAAGAAAAGTTTTTTCCTCAGAGCGGGGTTCTC 1620
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3763 CCGGCCATGGCAAGCCACAAGGACGATGAAGAAAAGTTTTTTCCTCAGAGCGGGGTTCTC 3822
Qy 1621 ATCTTTGGGAAGCAAGGCTCAGAGAAAACAAATGTGGACATTGAAAAGGTCATGATTACA 1680
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3823 ATCTTTGGGAAGCAAGGCTCAGAGAAAACAAATGTGGACATTGAAAAGGTCATGATTACA 3882
Qy 1681 GACGAAGAGGAAATCAGGACAACCAATCCCGTGGCTACGGAGCAGTATGGTTCTGTATCT 1740
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3883 GACGAAGAGGAAATCAGGACAACCAATCCCGTGGCTACGGAGCAGTATGGTTCTGTATCT 3942
Qy 1741 ACCAACCTCCAGAGAGGCAACAGACAAGCAGCTACCGCAGATGTCAACACACAAGGCGTT 1800
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3943 ACCAACCTCCAGAGAGGCAACAGACAAGCAGCTACCGCAGATGTCAACACACAAGGCGTT 4002
Qy 1801 CTTCCAGGCATGGTCTGGCAGGACAGAGATGTGTACCTTCAGGGGCCCATCTGGGCAAAG 1860
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4003 CTTCCAGGCATGGTCTGGCAGGACAGAGATGTGTACCTTCAGGGGCCCATCTGGGCAAAG 4062
Qy 1861 ATTCCACACACGGACGGACATTTTCACCCCTCTCCCCTCATGGGTGGATTCGGACTTAAA 1920
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4063 ATTCCACACACGGACGGACATTTTCACCCCTCTCCCCTCATGGGTGGATTCGGACTTAAA 4122
Qy 1921 CACCCTCCTCCACAGATTCTCATCAAGAACACCCCGGTACCTGCGAATCCTTCGACCACC 1980
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4123 CACCCTCCTCCACAGATTCTCATCAAGAACACCCCGGTACCTGCGAATCCTTCGACCACC 4182
Qy 1981 TTCAGTGCGGCAAAGTTTGCTTCCTTCATCACACAGTACTCCACGGGACAGGTCAGCGTG 2040
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4183 TTCAGTGCGGCAAAGTTTGCTTCCTTCATCACACAGTACTCCACGGGACAGGTCAGCGTG 4242
Qy 2041 GAGATCGAGTGGGAGCTGCAGAAGGAAAACAGCAAACGCTGGAATCCCGAAATTCAGTAC 2100
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4243 GAGATCGAGTGGGAGCTGCAGAAGGAAAACAGCAAACGCTGGAATCCCGAAATTCAGTAC 4302
Qy 2101 ACTTCCAACTACAACAAGTCTGTTAATGTGGACTTTACTGTGGACACTAATGGCGTGTAT 2160
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4303 ACTTCCAACTACAACAAGTCTGTTAATGTGGACTTTACTGTGGACACTAATGGCGTGTAT 4362
Qy 2161 TCAGAGCCTCGCCCCATTGGCACCAGATACCTGACTCGTAATCTGTAA 2208
||||||||||||||||||||||||||||||||||||||||||||||||
Db 4363 TCAGAGCCTCGCCCCATTGGCACCAGATACCTGACTCGTAATCTGTAA 4410
Alignment of instant SEQ ID NO: 27 with NCBI accession AF043303:
Query Match 99.5%; Score 615; Length 2208;
Best Local Similarity 100.0%;
Matches 615; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 4 CTGGAGACGCAGACTCAGTACCTGACCCCCAGCCTCTCGGACAGCCACCAGCAGCCCCCT 63
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 527 CTGGAGACGCAGACTCAGTACCTGACCCCCAGCCTCTCGGACAGCCACCAGCAGCCCCCT 586
Qy 64 CTGGTCTGGGAACTAATACGATGGCTACAGGCAGTGGCGCACCAATGGCAGACAATAACG 123
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 587 CTGGTCTGGGAACTAATACGATGGCTACAGGCAGTGGCGCACCAATGGCAGACAATAACG 646
Qy 124 AGGGCGCCGACGGAGTGGGTAATTCCTCGGGAAATTGGCATTGCGATTCCACATGGATGG 183
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 647 AGGGCGCCGACGGAGTGGGTAATTCCTCGGGAAATTGGCATTGCGATTCCACATGGATGG 706
Qy 184 GCGACAGAGTCATCACCACCAGCACCCGAACCTGGGCCCTGCCCACCTACAACAACCACC 243
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 707 GCGACAGAGTCATCACCACCAGCACCCGAACCTGGGCCCTGCCCACCTACAACAACCACC 766
Qy 244 TCTACAAACAAATTTCCAGCCAATCAGGAGCCTCGAACGACAATCACTACTTTGGCTACA 303
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 767 TCTACAAACAAATTTCCAGCCAATCAGGAGCCTCGAACGACAATCACTACTTTGGCTACA 826
Qy 304 GCACCCCTTGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTTTCACCACGTGACT 363
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 827 GCACCCCTTGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTTTCACCACGTGACT 886
Qy 364 GGCAAAGACTCATCAACAACAACTGGGGATTCCGACCCAAGAGACTCAACTTCAAGCTCT 423
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 887 GGCAAAGACTCATCAACAACAACTGGGGATTCCGACCCAAGAGACTCAACTTCAAGCTCT 946
Qy 424 TTAACATTCAAGTCAAAGAGGTCACGCAGAATGACGGTACGACGACGATTGCCAATAACC 483
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 947 TTAACATTCAAGTCAAAGAGGTCACGCAGAATGACGGTACGACGACGATTGCCAATAACC 1006
Qy 484 TTACCAGCACGGTTCAGGTGTTTACTGACTCGGAGTACCAGCTCCCGTACGTCCTCGGCT 543
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1007 TTACCAGCACGGTTCAGGTGTTTACTGACTCGGAGTACCAGCTCCCGTACGTCCTCGGCT 1066
Qy 544 CGGCGCATCAAGGATGCCTCCCGCCGTTCCCAGCAGACGTCTTCATGGTGCCACAGTATG 603
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1067 CGGCGCATCAAGGATGCCTCCCGCCGTTCCCAGCAGACGTCTTCATGGTGCCACAGTATG 1126
Qy 604 GATACCTCACCCTGA 618
|||||||||||||||
Db 1127 GATACCTCACCCTGA 1141
NCBI accession AAQ19311.1 teaches a sequence with 90% identity to SEQ ID NO: 50.
Alignment of instant SEQ ID NO: 50 with NCBI accession AAQ19311.1:
Query Match 100.0%; Score 799; Length 150;
Best Local Similarity 100.0%;
Matches 150; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MTTSGVPFGMTLRPTRSRLSRRTPYSRDRLPPFETETRATILEDHPLLPECNTLTMHNAW 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MTTSGVPFGMTLRPTRSRLSRRTPYSRDRLPPFETETRATILEDHPLLPECNTLTMHNAW 60
Qy 61 TSPSPPVKQPQVGQQPVAQQLDSDMNLSELPGEFINITDERLARQETVWNITPKNMSVTH 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 TSPSPPVKQPQVGQQPVAQQLDSDMNLSELPGEFINITDERLARQETVWNITPKNMSVTH 120
Qy 121 DMMLFKASRGERTVYSVCWEGGGRLNTRVL 150
||||||||||||||||||||||||||||||
Db 121 DMMLFKASRGERTVYSVCWEGGGRLNTRVL 150
At the time of filing it would have been obvious to one of ordinary skill in the art to combine the teachings of Braun et al into a single embodiment given that Braun exemplifies a method of producing AAV particles in a Brassica rapa plant comprising transforming said plants with construct comprising sequences which encode AAV particles and all the components necessary to assemble an AAV particle in planta and given that Braun exemplifies isolating and quantifying said AAV particles. One would have a reasonable expectation of success given the teachings of Braun et al.
At the time of filing, it would have been further obvious to one of ordinary skill in the art to further modify the teachings of Braun et al to include nucleic acid sequences encoding a REP protein, a CAP protein, an AAP protein, and an AD5 E4orf6 protein given the additional teachings of Braun et al and the teachings of Allen et al. One would have been motivated to include nucleic acid sequences encoding a REP protein, a CAP protein, an AAP protein, and an AD5 E4orf6 protein given that Braun teaches plans comprising VP1, VP2, VP3 (i.e. a CAP protein), AAP, Rep52 and Rep78 as well as an adenovirus helper plasmid and one of ordinary skill in the art would understand that these components are necessary for the production of functional AAV particles in a host. One would have been further motivated to replace the adenovirus helper plasmid of Braun et al with the minimal AD5 E4orf6 protein as taught by Allen et al given the explicit teachings of Allen regarding its benefits over a larger adenovirus helper plasmid
It would have been obvious to one of ordinary skill in the art to further modify the combined teachings of Braun et al and Allen et al to select the wild type AAV2 sequences disclosed by NCBI accession AF043303 and the E4orf6 sequence disclosed by AAQ19311.1 given the explicit suggestions of Braun et al that AAV2 sequences can be used to produce recombinant AAV in plant cells and given the teachings of Allen et al to use E4orf6.
Regarding claim 76, at the time of filing it would have been obvious to one of ordinary skill in the art to further modify the method of Braun et al to transform an Arabidopsis thaliana plant. One would be motivated to do with a reasonable expectation of the success given that Braun et al suggest using an Arabidopsis thaliana plant with their methods and the routine use of Arabidopsis thaliana in the art.
Regarding claim 77, at the time of filing it would have been obvious to one of ordinary skill in the art to further modify the method of Braun et al to include codon optimization of the nucleic acid sequences given that Braun et al teaches that the genes encoding the proteins required for the production of the recombinant viral vector can be codon optimized to improve expression in a plant cell. One would be motivated to do with a reasonable expectation of the success given the express teaching of Braun to use codon optimization and the routine nature of codon optimization for improved gene expression in plants.
Conclusion
No claims are allowed.
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/ALEKSANDAR RADOSAVLJEVIC/Examiner, Art Unit 1662
/BRENT T PAGE/Primary Examiner, Art Unit 1663
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