Production of herpes simplex viral protein
First Claim
1. A process for producing a polypeptide comprising the amino acid sequence of a Herpes Simplex Virus qD glycoprotein or a fragment of said amino acid sequence, wherein said fragment comprises at lest seven contiguous amino acids, said polypeptide containing at least one immunological and antigenic determinant of a Herpes Simplex Virus gD glycoprotein comprising:
- (a) culturing a unicellular organism containing a recombinant vector comprising a DNA sequence coding for a polypeptide comprising the amino acid sequence of a Herpes Simplex Virus gD glycoprotein or a fragment of said amino acid seqeunce, wherein said fragment comprises at least seven contiguous amino acids, said polypeptide containing at least one immunological and antigenic determinant of a Herpes Simplex Virus gD glycoprotein, and said recombinant vector capable of being replicated in said unicellular organism and said unicellular organism capable of producing said polypeptide; and
(b) isolating said polypeptide from the culture.
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Accused Products
Abstract
Methods and composition are provided for the cloning and expression of a Herpes Simplex Virus (HSV) glycoprotein (gD) gene in single-cell host organisms. Also described are methods for culturing these novel single-cell organisms to produce the HSV-gene product. The HSV gD-related polypeptide produced by the recombinant DNA techniques described herein may be formulated for use as an immunogen in a vaccine to protect against HSV-1 and HSV-2 infection.
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Citations
61 Claims
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1. A process for producing a polypeptide comprising the amino acid sequence of a Herpes Simplex Virus qD glycoprotein or a fragment of said amino acid sequence, wherein said fragment comprises at lest seven contiguous amino acids, said polypeptide containing at least one immunological and antigenic determinant of a Herpes Simplex Virus gD glycoprotein comprising:
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(a) culturing a unicellular organism containing a recombinant vector comprising a DNA sequence coding for a polypeptide comprising the amino acid sequence of a Herpes Simplex Virus gD glycoprotein or a fragment of said amino acid seqeunce, wherein said fragment comprises at least seven contiguous amino acids, said polypeptide containing at least one immunological and antigenic determinant of a Herpes Simplex Virus gD glycoprotein, and said recombinant vector capable of being replicated in said unicellular organism and said unicellular organism capable of producing said polypeptide; and (b) isolating said polypeptide from the culture. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 45)
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15. A process for preparing a unicellular organism having a DNA sequence coding for a polypeptide comprising the amino acid sequence of a Herpes Simplex Virus gD glycoprotein or a fragment of said amino acid sequence, wherein said fragment comprises at least seven contiguous amino acids, said polypeptide containing at least one immunolgocial and antigenic determinant of a Herpes Simplex Virus gD glycoprotein, comprising:
- introducing a recombinant vector into a unicellar orgainism, said recombinant vector comprising a DNA sequence coding for a polypeptide comprising the amino acid sequence of a Herpes Simplex Virus gD glycoprotein or a fragment of said amino acid sequence, wherein said fragment comprises at least seven contiguous amino acids, said polypeptide containing at least one immunological and antigenic determinant of a Herpes Simplex Virus gD glycoprotein, and said recombinant vector capable of being replicated in said unicellar organism and said unicellar organism capable of producing said polypeptide.
- View Dependent Claims (16, 17)
- 18. A purified DNA sequence coding for a polypeptide comprising the amino acid sequence of a Herpes Simplex virus gD glycoprotein or a fragment of said amino acid sequence, wherein said fragment comprises at least seven contiguous amino acids, said polypeptide containing at least one immunological and antigenic determinant of a Herpes Simplex Virus gD glycoprotein, in which such DNA sequence is free of Herpes Simplex Virus-coded translation stop signals in phase and 5'"'"', to the triplet signal coding for the first amino acid of said polypeptide.
- 22. A unicellular organism containing an exogenous DNA sequence coding for a polypeptide comprising the amino acid sequence of a Herpes Simplex Virus gD glycoprotein or a fragment of said amino acid sequence, wherein said fragment comprises at least seven contiguous amino acids, said polypeptide containing at least one immunological and antigenic determinant of a Herpes Simplex Virus gD glycoprotein, wherein the unicellular orgainism is capable of expressing the DNA sequence to produce said polypeptide.
- 23. A unicellular organism containing a recombinant vector comprising a DNA sequence coding for polypeptide comprsiing the amino acid sequence of a Herpes Simplex Virus gD glycoprotein or a fragment of said amino acid sequence, wherein said fragment comprises at least seven contiguous amino acids, said polypeptide containing at least one immunological and antigeninc determinant of a Herpes Simplex Virus gD glycoprotein, wherein said recombinant vector is capable of being replicated in said unicellular organism and said unicellular orgainism is capable of expressing the DNA sequence to produce said polypeptide.
- 32. A recombinant vector comprising (a) a DNA sequence coding for a polypeptide comprising the amino acid sequence of a Herpes Simplex Virus gD glycoprotein or a fragment of said amino acid sequence, wherein said fragment comprises at least seven contiguous amino acids, said polypeptide containing at least one immunological and antigenic determinant of a Herpes Simplex Virus gD glycoprotein and (b) DNA from a vector derived from a plasmid, virus or cell which does not exchange genetic information with Herpes Simplex Virus in nature, wherien the recombinant vector is capable of being replicated in a compatible host cell and capable of directing expression of the DNA sequence in said host cell to produce said polypeptide.
- 37. A eucaryotic cell containing an exgenous DNA sequence coding for a polypeptide comprising the amino acid sequence of a Herpes Simplex Virus gD glycoprotein or a fragment of said amino acid sequence, wherein said fragment comprises at least seven contiguous amino acids, said polypeptide containing at least one immunological and antigenic determinant of a Herpes Simplex Virus gD glycoprotein, wherein the eucaryotic cell is capable of expressing the DNA sequence to produce said polypeptide and does not contain the entirety of the Herpes Simplex Virus genetic information.
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38. A eucaryotic cell containin an exogenous DNa sequence coding for a polypeptide comprising the amino acid sequence of a Herpes Simplex Virus gD glycoprotein or a fragement of said amino acid sequence, wherein said fragment comprises at least seven contiguous amino acids, said polypeptide containing at least one immunological and antigenic determinant of a herpes Simplex Virus gD glycoprotein, wherein the eucaryotic cell is capable of expressing the DNA sequence to produce said polypeptide and is not a natural host for Herpes Simplex Virus.
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41. The recombinant vector pEH51.
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42. The recombinant vector pEH82.
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43. A unicellular organism containing the recombinant vector pEH51.
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44. A procarbyotic cell containing an exogenous DNA sequence coding for a polypeptide comprising the amino acid sequence of a Herpes Simplex Virus gD glycoprotein or a fragment of said amino acid sequence, wherein said fragment comprises at least seven contiguous amino acids, said polypeptide containing at least one immunological and antigenic determinant of a Herpes Simplex Virus gD glycoprotein, wherein the procaryotic cell is capable of expressing the DNA sequence to produce said polypeptide.
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46. A unicellular organism containing the exogenous DNA sequence:
- GGG GGG ACT GCC GCC AGG TTG GGG GCC GTG ATT TTG TTT GTC GTC ATA GTG GGC CTC CAT GGG GTC CGC GGC AAA TAT GCC TTG GCG GAT GCC TCT CTC AAG ATG GCC GAC CCC AAT CGC TTT CGC GGC AAA GAC CTT CCG GTC CTG GAC CAG CTG ACC GAC CCT CCG GGG GTC CGG CGC GTG TAC CAC ATC CAG GCG GGC CTA CCG GAC CCG TTC CAG CCC CCC AGC CTC CCG ATC ACG GTT TAC TAC GCC GTG TTG GAG CGC GCC TGC CGC AGC GTG CTC CTA AAC GCA CCG TCG GAG GCG CCC CAG ATT GTC CGC GGG GCC TCC GAA GAC GTC CGG AAA CAA CCC TAC AAC CTG ACC ATC GCT TGG TTT CGG ATG GGA GGC AAC TGT GCT ATC CCC ATC ACG GTC ACG GAG TAC ACC GAA TGC TCC TAC AAC AAG TCT CTG GGG GCC TGT CCC ATC CGA ACG CAG CCC CGC TGG AAC TAC TAT GAC AGC TTC AGC GCC GTC AGC GAG GAT AAC CTG GGG TTC CTG ATG CAC GCC CCC GCG TTT GAG ACC GCC GGC ACG TAC CTG CGG CTC GTG AAG ATA AAC GAC TGG ACG GAG ATT ACA CAG TTT ATC CTG GAG CAC CGA GCC AAG GGC TCC TGT AAG TAC GCC CTC CCG CTG CGC ATC CCC CCG TCA GCC TGC CTC TCC CCC CAG GCC TAC CAG CAG GGG GTG ACG GTG GAC AGC ATC GGG ATG CTG CCC CGC TTC ATC CCC GAG AAC CAG CGC ACC GTC GCC GTA TAC AGC TTG AAG ATC GCC GGG TGG CAC GGG CCC AAG GCC CCA TAC ACG AGC ACC CTG CTG CCC CCG GAG CTG TCC GAG ACC CCC AAC GCC ACG CAG CCA GAA CTC GCC CCG GAA GAC CCC GAG GAT TCG GCC CTC TTG GAG GAC CCC GTG GGG ACG GTG GCG CCG CAA ATC CCA CCA AAC TGG CAC ATC CCG TCG ATC CAG GAC GCC GCG ACG CCT TAC CAT CCC CCG GCC ACC CCG AAC AAC ATG GGG CTG ATC GCC GGC CGG GTG GGC GGC AGT CTC CTG GCA GCC CTG GTC ATT TGC GGA ATT GTG TAC TGG ATG CAC CGC CGC ACT CGG AAA GCC CCA AAG CGC ATA CGC CTC CCC CAC ATC CGG GAA GAC GAC CAG CCG TCC TCG CAC CAG CCC TTG TTT TAC, or a subsequence thereof, which subsequence comprises at least 21 continguous nucleotides of said sequence and codes on expression for a polypeptide having at least one immunological and antigenic determinant of a Herpoes Simplex Virus gD glycoprotein, wherein said polypeptide comprises at least seven contiguous amino acids and said unicellular organism is capable of expressing the DNA sequence to produce said polypeptide.
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47. A recombinant vector comprising the DNA sequennce:
- GGG GGG ACT GCC GCC AGG TTG GGG GCC GTG ATT TTG TTT GTC GTC ATA GTG GGC CTC CAT GGG GTC CGC GGC AAA TAT GCC TTG GCG GAT GCC TCT CTC AAG ATG GCC GAC CCC AAT CGC TTT CGC GGC AAA GAC CTT CCG GTG CTG GAC CAG CTG ACC GAC CCT CCG GGG GTC CGG CGC GTG TAC CAC ATC CAG GCG GGC CTA CCG GAC CCG TTC CAG CCC CCC AGC CTC CCG ATC ACG GTT TAC TAC GCC GTG TTG GAG CGC GCC TGC CGC AGC GTG CTC CTA AAC GCA CCG TCG GAG GCG CCC CAG ATT GTC CGC GGG GCC TCC GAA GAC GTC CGG AAA CAA CCC TAC AAC CTG ACC ATC GCT TGG TTT CGG ATG GGA GGC AAC TGT GCT ATC CCC ATC ACG GTC ACG GAG TAC ACC GAA TGC TCC TAC AAC AAG TCT CTG GGG GCC TGT CCC ATC CGA ACG CAG CCC CGC TGG AAC TAC TAT GAC AGC TTC AGC GCC GTC AGC GAG GAT AAC CTG GGG TTC CTG ATG CAC GCC CCC GCG TTT GAG ACC GCC GGC ACG TAC CTG CGG CTC GTG AAG ATA AAC GAC TGG ACG GAG ATT ACA CAG TTT ATC CTG GAG CAC CGA GCC AAG GGC TCC TGT AAG TAC GCC CTC CCG CTG CGC ATC CCC CCG TCA GCC TGC CTC TCC CCC CAG GCC TAC CAG CAG GGG GTG ACG GTG GAC AGC ATC GGG ATG CTG CCC CGC TTC ATC CCC GAG AAC CAG CGC ACC GTC GCC GTA TAC AGC TTG AAG ATC GCC GGG TGG CAC GGG CCC AAG GCC CCA TAC ACG AGC ACC CTG CTG CCC CCG GAG CTG TCC GAG ACC CCC AAC GCC ACG CAG CCA GAA CTC GCC CCG GAA GAC CCC GAG GAT TCG GCC CTC TTG GAG GAC CCC GTG GGG ACG GTG GCG CCG CAA ATC CCA CCA AAC TGG CAC ATC CCG TCG ATC CAG GAC GCC GCG ACG CCT TAC CAT CCC CCG GCC ACC CCG AAC AAC ATG GGG CTG ATC GCC GGC GCG GTG GGC GGC AGT CTC CTG GCA GCC CTG GTC ATT TGC GGA ATT GTG TAC TGG ATG CAC CGC CGC ACT CGG AAA GCC CCA AAG CGC ATA CGC CTC CCC CAC ATC CGG GAA GAC GAC CAG CCG TCC TCG CAC CAG CCC TTG TTT TAC, or a subsequence thereof which subsequence comprises at least 21 contiguous nucleotide of said sequence and codes on expression for a polypeptide having at least one immunological and natigenic determinant of a Herpes Simplex Virus gD glycoprotein, wherein said polypeptide comprises at least seven contiguous amino acids and said recombinant vector is capable of being replicated in a compatible host cell and capable of directing expression of the DNA sequence in said host cell to produce said polypeptide.
- View Dependent Claims (48, 49, 50)
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51. A unicellular organism containing a recombinanat vector comprising the DNA sequence:
- GGG GGG ACT GCC GCC AGG TTG GGG GCC GTG ATT TTG TTT GTC GTC ATA GTG GGC CTC CAT GGG GTC CGC GGC AAA TAT GCC TTG GCG GAT GCC TCT CTC AAG ATG GCC GAC CCC AAT CGC TTT CGC GGC AAA GAC CTT CCG GTC CTG GAC CAG CTG ACC GAC CCT CCG GGG GTC CGG CGC GTG TAC CAC ATC CAG GCG GGC CTA CCG GAC CCG TTC CAG CCC CCC AGC CTC CCG ATC ACG GTT TAC TAC GCC GTG TTG GAG CGC GCC TGC CGC AGC GTG CTC CTA AAC GCA CCG TCG GAG GCG CCC CAG ATT GTC CGC GGG GCC TCC GAA GAC GTC CGG AAA CAA CCC TAC ACC CTG ACC ATC GCT TGG TTT CGG ATG GGA GGC AAC TGT GCT ATC CCC ATC ACG GTC ACG GAG TAC ACC GAA TGC TCC TAC AAC AAG TCT CTG GGG GCC TGT CCC ATC CGA ACG CAG CCC CGC TGG AAC TAC TAT GAC AGC TTC AGC GCC GTC AGC GAG GAT AAC CTG GGG TTC CTG ATG CAC GCC CCC GCG TTT GAG ACC GCC GGC ACG TAC CTG CGG CTC GTG AAG ATA AAC GAC TGG ACG GAG ATT ACA CAG TTT ATC CTG GAG CAC CGA GCC AAG GGC TCC TGT AAG TAC GCC CTC CCG CTG CGC ATC CCC CCG TCA GCC TGC CTC TCC CCC CAG GCC TAC CAG CAG GGG GTG ACG GTG GAC AGC ATC GGG ATG CTG CCC CGC TTC ATC CCC GAG AAC CAG CGC ACC GTC GCC GTA TAC AGC TTG AAG ATC GCC GGG TGG CAC GGG CCC AAG GCC CCA TAC ACG AGC ACC CTG CTG CCC CCG GAG CTG TCC GAG ACC CCC AAC GCC ACG CAG CCA GAA CTC GCC CCG GAA GAC CCC GAG GAT TCG GCC CTC TTG GAG GAC CCC GTG GGG ACG GTG CCG CCG CAA ATC CCA CCA AAC TGG CAC ATC CCG TCG ATC CAG GAC GCC GCG ACG CCT TAC CAT CCC CCG GCC ACC CCG AAC AAC ATG GGG CTG ATC GCC GGC GCG GTG GGC GGC AGT CTC CTG GCA GCC CTC GTC ATT TGC GGA ATT GTG TAC TGG ATG CAC CGC CGC ACT CGG AAA GCC CCA AAG CGC ATA CGC CTC CCC CAC ATC CGG GAA GAC GAC CAG CCG TCC TCG CAC CAG CCC TTG TTT TAC, or a subsequence thereof, which subsequence comprises at least 21 contiguous nuleotides of said sequence and codes on expression for a polypeptide having at least one immunological and antigenic determinant of a Herpes Simplex Virus gD glycoprotein, wherein said polypeptide comprises at least seven contiguous amino acids and said recombinant vector is capable of being replicated in said unicellular organism and said unicellular orgainsim is capable of expressing the DNA sequence to produce said polypeptide.
- View Dependent Claims (52, 53, 54)
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55. An Escherichia coli bacterium as deposited with the ATCC and assigned accession No. 39,159.
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56. An Escherichia coli bacterium as deposited with the ATCC and assigned aaccession No. 39,160.
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57. An Escherichia coli bacterium containing the exogenous DNA sequence:
- GGG GGG ACT GCC GCC AGG TTG GGG GCC GTG ATT TTG TTT GTC GTC ATA GTG GGC CTC CAT GGG GTC CGC GGC AAA TAT GCC TTG GCG GAT GCC TCT CTC AAG ATG GCC GAC CCC AAT CGC TTT CGC GGC AAA GAC CTT CCG GTC CTG GAC CAG CTG ACC GAC CCT CCG GGG GTC CGG CGC GTG TAC CAC ATC CAG GCG GGC CTA CCG GAC CCG TTC CAG CCC CCC AGC CTC CCG ATC ACG GTT TAC TAC GCC GTG TTG GAG CGC GCC TGC CGC AGC GTG CTC CTA AAC GCA CCG TCG GAG GCG CCC CAG ATT GTC CGC GGG GCC TCC GAA GAC GTC CGG AAA CAA CCC TAC AAC CTG ACC ATC GCT TGG TTT CGG ATG GGA GGC AAC TGT GCT ATC CCC ATC ACG GTC ACG GAG TAC ACC GAA TGC TCC TAC AAC AAG TCT CTG GGG GCC TGT CCC ATC CGA ACG CAG CCC CGC TGG AAC TAC TAT GAC AGC TTC AGC GCC GTC AGC GAG GAT AAC CTG GGG TTC CTG ATG CAC GCC CCC GCG TTT GAG ACC GCC GGC ACG TAC CTG CGG CTC GTC AAG ATA AAC GAC TGG ACG GAG ATT ACA CAG TTT ATC CTG GAG CAC CGA GCC AAG GGC TCC TGT AAG TAC GCC CTC CCG CTG CGC ATC CCC CCG TCA GCC TGC CTC TCC CCC CAG GCC TAC CAG CAG GGG GTG ACG GTG GAC AGC ATC GGG ATG CTG CCC CGC TTC ATC CCC GAG AAC CAG CGC ACC GTC GCC GTA TAC AGC TTG AAG ATC GCC GGG TGG CAC GGG CCC AAG GCC CCA TAC ACG AGC ACC CTG CTG CCC CCG GAG CTG TCC GAG ACC CCC AAC GCC ACG CAG CCA GAA CTC GCC CCG GAA GAC CCC GAG GAT TCG GCC CTC TTG GAG GAC CCC GTG GGG ACG GTG GCG CCG CAA ATC CCA CCA AAC TGG CAC ATC CCG TCG ATC CAG GAC GCC GCG ACG CCT TAC CAT CCC CCG GCC ACC CCG AAC AAC ATG GGG CTG ATC GCC GGC GCG GTG GGC GGC AGT CTC CTG GCA GCC CTG GTC ATT TGC GGA ATT GTG TAC TGG ATG CAC CGC CGC ACT CGG AAA GCC CCA AAG CGC ATA CGC CTC CCC CAC ATC CGG GAA GAC GAC CAG CCG TCC TCG CAC CAG CCC TTG TTT TAC, or a subsequence thereof, which subsequence comprises at least 21 contiguous nucleotides of said sequence and codes on expression for a polypeptide having at least one immunological and antigenic determinant of a Herpes Simplex Virus gD glycoprotein, wherein said polypeptide comprises at least seven contiguous amino acids and said bacterium is capable of expressing the DNA sequence to produce said polypeptide.
- 58. An Escherichia coli bacterium containing an exogenous DNA sequence coding for a polypeptide comprising the amino acid sequence of a Herpes Simplex Virus gD glycoprotein or a fragment of said amino acid sequence, wherein said fragment comprises at least seven contignuous amino acids, said polypeptide containing at least one immunological and antigenic determinanat of a Herpes Simplex Virus gD glycoprotein, and wherein the bacterium is capable of expressing the DNa sequence to produce said polypeptide.
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59. An Escherichia coli bacterium containing a recombinant vector comprising a DNA sequence coding for a polypeptide comprising the amino acid sequence of a Herpes Simplex Virus gD glycoprotein or a fragement of said amino acid sequence, wherein said fragment comprises at least seven contiguous amino acids, said polypeptide containing at least one immunolgocal and antigenic determinanat of a herpes Simplex Virus gD glycoprotein, in which such DNa sequence is connected in phase to a second DNA sequence coding for a protein, and the recombinant vector is capable of being replicated in said bacterium and said bacterium is capable of expressing the DNA sequence to produce said polypeptide.
Specification