Production of herpes simplex viral protein

• US 4,818,694 A
• Filed: 07/20/1982
• Issued: 04/04/1989
• Est. Priority Date: 07/20/1982
• Status: Expired due to Term

• Current Assignee: American Cyanamid Co (Pfizer Inc.)
• Original Assignee: American Cyanamid Co (Pfizer Inc.)
• Inventors: Weis, John W., Enquist, Lynn W., Watson, Roger J.
• Primary Examiner(s): Martinell, James
• Application Number:
  US06/400,028
• Time in Patent Office: 2,450 Days
• Field of Search: 435/68, 435/70, 435/91, 435/172, 435/253, 435/317, 435/317.1, 435/320, 435/255, 435/256, 435/240.1, 435/240.2, 435/71, 435/172.3, 435/172.1, 435/243, 536/27, 935/11, 935/12, 935/52, 935/66-75, 935/28, 935/29, 935/31, 935/32
• US Class Current: 435/69.3
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: A61K 39/245 Herpetoviridae, e.g. herpes simplex virus
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/40 containing a tag for immunodetection, or an epitope for immunisation
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins
• CPC Class Codes: C12N 15/70 Vectors or expression systems specially adapted for E. coli
• CPC Class Codes: C12N 2710/16622 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2710/16634 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: Y10S 930/224 Herpes related

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.

Claims

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.

2. The process according to claim 1, wherein the recombinant vector was introduced into the unicellular organism by transformation.

3. The process according to claim 1, wherein the recombinant vector was introduced into the unicellular organism by transduction.

4. The process according to claim 1, wherein the recombinant vector was introduced into the unicellular organism by transfection.

5. The process according to claim 1, wherein the information contained in the DNA sequence was obtained from a Herpes Simplex Virus genome.

6. The process according to claim 5, wherein the information contained in the DNA sequence was obtained from Herpes Simplex Virus type 1.

7. The process according to claim 1, wherein the information contained in the DNA sequence was obtained by isolating mRNA coding for a Herpes Simplex Virus gD glycoprotein and using reverse transcriptase to construct said DNA sequence having one strand complementary to the isolated mRNA.

8. The process according to claim 1, wherein the DNA sequence was obtained from a DNA vector containing a Herpes Simplex virus DNA sequence.

9. The process according to claim 1, wherein the DNA sequence codes for the gD glycoprotein of Herpes Simplex Virus type 1.

10. The process according to claim 1, wherein the unicellular organism is a eucaryotic organism.

11. The process according to claim 1, wherein the unicellular organism is a procaryotic organism.

12. The process according to claim 11, wherein the procaryotic organism is Escherichia coli.

13. The process according to claim 12, wherein the Escherichia coli has ATCC accession No. 39,159.

14. The process according to claim 12, wherein the Escherichia coli has ATCC accession No. 39,160.

45. The process of claim 1, wherein said unicellular organism is a yeast.

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.

16. The process of claim 17, wherein the Herpes Simplex Virus is Herpes Simplex Virus Type 1.

17. The process of claim 15, wherein at least one determinant is common to Herpes Simplex Virus Type 1 and Type 2.

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.

19. The purified DNA sequence of claim 18, in which the determinnat is specific to Herpes Simplex Virus Type 1.

20. The purified DNA sequence of claim 18, in which the determinant is common to Herpes Simplex Virus Type 1 and Type 2.

21. The purified DNA sequence of claim 18, in which the sequence comprises 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 TTTG 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 CGC 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 CTC 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 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.

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.

24. The unicellular organism of claim 22 or 23, wherein the Herpes Simplex Virus is Herpes Simplex Virus Type 1.

25. The unicellular organism of claim 22 or 23, wherein at least one determinanat is common to Herpes Simplex Virus Type 1 and Type 2.

26. The unicellular organism of claim 22 or 23, wherein the unicellular organism is a yeast.

27. The unicellar organism of claim 24, wherein the unicellar organism is a yeast.

28. The unicellular organism of claim 25, wherein the unicellular organism is a yeast.

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.

29. The unicellular organism of claim 23, in which the DNA sequence in such recombinant vector is under the control of a transcription promoter and translation initiation signal.

30. The unicellular organism of claim 23, in which such recombinant vector further comprises the PBR322 replication origin.

31. The unicellar organism of claim 23, in which the DNA seqeunce in such recombinant vector is connected in phase to a second DNA sequence coding for a protein.

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.

33. The recombinant vector of claim 32, further comprising the PBR322 replication origin.

34. The recombinant vector of claim 32, in which the DNA sequence is connected in phase to a second DNA sequence coding for a protein.

35. The recombinant vector of claim 32, wherein the Herpes Simplex Virus is Herpes Simplex Virus Type 1.

36. The recombinant vector of claim 32, wherein at least one determinant is common to Herpes Simplex Virus Type 1 and Type 2.

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.

39. The eucaryotic cell of claim 37 or 38, wherein the Herpes Simplex Virus is Herpes Simplex Virus Type 1.

40. The eucaryotic cell of claim 37 or 38, wherein at least one determinant is common to Herpes Simplex Virus Type 1 and Type 2.

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.

41. The recombinant vector pEH51.

42. The recombinant vector pEH82.

43. A unicellular organism containing the recombinant vector pEH51.

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.

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.

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.

48. The recombinant vector of claim 47, wherein said DNA sequence is under the control of a transcription promoter and at translation initiation signal.

49. The recombinant vector of claim 47, further comprising the pBR322 replication origin.

50. The recombinant vector of claim 47, wherein said DNA sequence is connected in phases to a second DNA sequence coding for a protein.

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.

52. The unicellular organism of claim 51, wherein said DNA sequence is under the control of a transcription promoter and a translation initiation signal.

53. The unicellular organism of claim 51, wherein said recombinant vector comprises the pBR322 replication origin.

54. The unicellular organism of claim 51, wherein said DNA sequence is connected in phase to a second DNA sequence coding for a protein.

55. An Escherichia coli bacterium as deposited with the ATCC and assigned accession No. 39,159.

56. An Escherichia coli bacterium as deposited with the ATCC and assigned aaccession No. 39,160.

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.

60. The bacterium of claim 58 or 59, wherein the Herpes Simplex Virus is Herpes Simplex Virus Type 1.

61. The bacterium of claim 58 or 59, wherein at least one determinant is common to Herpes Simplex Virus Type 1 and Type 2.

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.