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Expression of recombinant glyoprotein B from herpes simplex virus
| Details |
Inventors: Burke, Rae L.; Pachl, Carol; Valenzuela, Pablo D. T.;
Assignee: Chiron Corporation (Emeryville, CA)
Primary Examiner: Chambers; Jasemine C.
Assistant Examiner:
Attorney, Agent or Firm: McClung; Barbara G., Robins; Roberta L.
The glycoprotein B ("gB") and functional fragments thereof are provided by recombinant DNA technology. Oligonucleotide sequences are provided coding the glycoprotein, its precursor and fragments thereof. Methods and compositions are disclosed for the production of the glycoprotein and functional fragments thereof as well as oligonucleotide sequences, which may be used for probes or other applications, and particularly may be used for vaccines. The following E. coli HB101 strains were deposited at the A.T.C.C. on April 4, 1984, where the plasmid indicates the plasmid employed to transform the strain; pHS112; pHS114 (gB1 mammalian); pHS127A (gB1 yeast); pHS203; and pHS206 (gB2), and assigned Accession Nos. 39649-39653, respectively, where the nature of the plasmid is indicated in parentheses and the number refers back to the number employed in the Experimental section. |
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DETAILED DESCRIPTION The nucleotide sequence and the amino acid sequence derived therefrom are provided for Herpes Simplex Virus ("HSV") glycoprotein B ("gB") for use in expression of the glycoprotein, synthesis of constructs for expression, production of the glycoprotein, use of the glycoprotein in vaccines, and use of the nucleotide fragments in the aforementioned constructions and as probes.
Particularly, DNA was identified as coding for gB and was manipulated with expression vectors for expression in eukaryotic cells.
The precursor and mature gB proteins and portions thereof are provided free of HSV genomic DNA.
Vaccines containing gB protein as well as methods for preparing and using the vaccines are also provided.
Accordingly, one aspect of the invention provides a DNA construct which contains an oligonucleotide sequence substantially free of natural flanking regions, which encodes a polypeptide similar to glycoprotein B of HSV or its functional fragments.
The DNA construct also contains transcriptional and translational regulatory sequences flanking the oligonucleotide sequence; at least one of these regulatory sequences is other than an HSV regulatory sequence.
In addition, the construct contains sequences which allow for its expression in a eukaryotic system.
Another aspect of the invention provides a eukaryotic host harboring the expression construct as well as a system which allows for replication of the construct, which is joined to the construct.
Another aspect of the invention provides a vaccine which contains a recombinant polypeptide which is immunologically cross reactive with glycoprotein B of HSV.
The polypeptide is present in an amount effective to produce an immune response in a mammalian host.
The polypeptide is synthesized via the expression of the DNA construct described above.
Another aspect of the invention provides a method for preparing vaccines against herpes simplex virus.
In this method an immunologically active polypeptide which is cross-reactive with glycoprotein B of HSV is synthesized in a eukaryotic host
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Gene Therapy 
