Recombinant proteins containing repeating units
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IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12N-015/64
C12N-015/00
C12N-015/09
C12N-015/63
C12P-019/34
C12P-019/00
출원번호
US-0804733
(2001-03-13)
발명자
/ 주소
Wang,Qi
Guan,Zhonghon
Baggot,Brendan O.
Hadfield,Kristen
Zhao,Jianmin
Edwards,Janice
출원인 / 주소
Monsanto Technology LLC
대리인 / 주소
Fulbright &
인용정보
피인용 횟수 :
1인용 특허 :
24
초록▼
Methods for the production of recombinant proteins containing repeating units are disclosed. Also disclosed are methods for the production of degenerate polynucleotides encoding said recombinant proteins. In addition, polypeptides and polynucleotides produced by the methods of current invention are
Methods for the production of recombinant proteins containing repeating units are disclosed. Also disclosed are methods for the production of degenerate polynucleotides encoding said recombinant proteins. In addition, polypeptides and polynucleotides produced by the methods of current invention are also disclosed.
대표청구항▼
What is claimed is: 1. A method for producing a recombinant protein containing repeating units comprising: (a) providing a first pool of polynucleotides comprising codons, said pool of polynucleotides comprising at least two tandem repeats of sequences encoding a portion of said recombinant protein
What is claimed is: 1. A method for producing a recombinant protein containing repeating units comprising: (a) providing a first pool of polynucleotides comprising codons, said pool of polynucleotides comprising at least two tandem repeats of sequences encoding a portion of said recombinant protein, wherein said tandem repeats contain degenerate nucleotide sequences encoding for a portion of said recombinant protein in accordance with the degeneracy of the genetic code; (b) providing a second pool of polynucleotides, at least some of which are complementary to the polynucleotides in said first pool of polynucleotides; (c) combining said first pool of polynucleotides and said second pool of polynucleotides under conditions whereby the polynucleotides will anneal; (d) extending the 3' ends of said annealed polynucleotides under conditions wherein said annealed polynucleotides act as primers for their complements; (e) denaturing the extended polynucleotides; (f) repeating steps (c)-(e) at least once, whereby the products of step (e) provide the polynucleotides for annealing in step (c) of the next cycle; (g) if necessary, adding one or more linker oligonucleotides to the end of the products of (f), said linker oligonucleotides containing at least one restriction enzyme cleavage site; (h) inserting the products of (f) or (g) into a suitable vector; (i) introducing said vector into a suitable host cell; and (j) maintaining said host cell under conditions allowing for expression of said recombinant protein. 2. The method of claim 1, wherein said tandem repeats are separated by no more than nine nucleotides. 3. The method of claim 1, further comprising cleaving said recombinant protein between said repeating units to produce non-repeating peptides. 4. The method of claim 3, further comprising cleaving said polynucleotides after step (f). 5. The method of claim 1, wherein at least 25% of the codons in said polynucleotides encode a desired amino acid. 6. The method of claim 1, wherein at least 50% of the codons in said polynucleotides encode a desired amino acid. 7. The method of claim 1, wherein at least 75% of the codons in said polynucleotides encode a desired amino acid. 8. The method of claim 1, wherein at least 90% of the codons in said palynucleotides encode a desired amino acid. 9. The method of claim 1, wherein said tandem repeats encode at least one sequence selected from the group consisting of LKPNM (SEQ ID NO:1), KPNM (SEQ ID NO:2), VVYP (SEQ ID NO:3), KPN, DKP, YKP, EKP, DAP, EAP, HPP, VPP, LK, PN and NM. 10. The method of claim 1, further comprising introducing a second vector into said host cell, said second vector containing a nucleotide sequence encoding an enzyme capable of cleaving said recombinant protein between said repeating units. 11. The method of claim 10, wherein said second vector further comprises a tissue or organelle specific promoter such that expression of said enzyme is restricted to a tissue or organelle different from the tissue or organelle expressing said recombinant protein. 12. The method of claim 10, wherein said second vector further comprises a plastid targeting sequence. 13. The method of claim 1, wherein said vector further comprises an expression cassette. 14. The method of claim 13, wherein said expression cassette comprises at least one promoter chosen from the group consisting of a tissue specific promoter, an inducible promoter, a constitutive promoter, a developmentally regulated promoter, an organelle specific promoter, a seed specific promoter and a plastid specific promoter. 15. The method of claim 13, wherein said expression cassette further comprises at least one plastid targeting sequence. 16. The method of claim 13, wherein said expression cassette further comprises at least one secretion sequence. 17. The method of claim 13, wherein said expression cassette further comprises an additional nucleotide sequence encoding an enzyme capable of cleaving said recombinant protein between said repeating units. 18. The method of claim 17, wherein said additional nucleotide sequence is under the control of a separate promoter. 19. The method of claim 1, wherein said host cell is selected from the group consisting of bacterial cells, yeast cells, insect cells and animal cells. 20. The method of claim 1, wherein said host cell is a plant cell.
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이 특허에 인용된 특허 (24)
Maugh Kathy J. (Walnut CA) Anderson David M. (Rockville MD), Escherichia coli expression vector encoding bioadhesive precursor protein analogs comprising three to twenty r.
Ferrari Franco A. (La Jolla CA) Richardson Charles (San Diego CA) Chambers James (San Diego CA) Causey Stuart C. (Del Mar CA) Pollock Thomas J. (San Diego CA) Capello Joseph (San Diego CA) Crissman J, Construction of synthetic DNA and its use in large polypeptide synthesis.
Ferrari Franco A. ; Richardson Charles ; Chambers James ; Causey Stuart ; Pollock Thomas J. ; Cappello Joseph ; Crissman John W., Peptides comprising repetitive units of amino acids and DNA sequences encoding the same.
Mullis Kary B. (La Jolla CA) Erlich Henry A. (Oakland CA) Arnheim Norman (Woodland Hills CA) Horn Glenn T. (Emeryville CA) Saiki Randall K. (Richmond CA) Scharf Stephen J. (Berkeley CA), Process for amplifying, detecting, and/or cloning nucleic acid sequences.
Mullis Kary B. (Kensington CA) Erlich Henry A. (Oakland CA) Arnheim Norman (Woodland Hills CA) Horn Glenn T. (Emeryville CA) Saiki Randall K. (Richmond CA) Scharf Stephen J. (Berkeley CA), Process for amplifying, detecting, and/or-cloning nucleic acid sequences.
Hopp Thomas P. (Seattle WA) Bektesh Susan L. (Seattle WA) Conlon ; III Paul J. (Seattle WA) March Carl J. (Seattle WA), Synthesis of protein with an identification peptide.
Hopp Thomas P. (Seattle WA) Bektesh Susan L. (Seattle WA) Conlon ; III Paul J. (Seattle WA) March Carl J. (Seattle WA), Synthesis of protein with an identification peptide, and hybrid polypeptide incorporating same.
Prakash, Indra; DuBois, Grant E., High-potency sweetener composition with rubisco protein, rubiscolin, rubiscolin derivatives, ace inhibitory peptides, and combinations thereof, and compositions sweetened therewith.
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