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Vector encoding human cytomegalovirus glycoprotein; plasmid pSP65IT construction; HXLF1 and HXLF2 gene cloning and expression; recombinant vaccine construction; application in diagnosis and therapy
Patent Report Antigenic polypeptides of Taenia ovis; D N A sequence, gene cloning and expression in Escherichia coil, yeast, mammal, insect cell culture; application to vaccine production Min. Agr. Fish. N.Z.; Coopers-Anita. Health; Univ. Melbourne UK 2219 590; 13 December 1989 A purified antigenic polypeptide or an immunologically active fragment is claimed, the polypeptide having a mol.wt of 47000-52000 calculated by SDS-PAGE and is capable of generating a protective immunological response to Taenia ovis in a ruminant. Also claimed is a DNA molecule comprising the DNA sequence which encodes the polypeptide or fragment. Also new are recombinant expression vectors containing the DNA molecule and host cells transformed with the vectors and capable of expressing the T. ovis polypeptide. The vectors plasmid pSjl0 Bam7Stop7-45W, plasmid pGEXI-45W and plasmid pGEX-2-45W are specifically claimed. The host cell transformed with the plasmids are Escherichia coli strain JM 101 or JM109, a mammal, yeast or insect cell. The polypeptide may be isolated from T. ovis eggs or by recombinant DNA techniques. The DNA is obtained using RNA isolated from T. ovis eggs. The polypeptide can be used in a ruminant vaccine against T. ovis infection and also in the vaccination of ruminants against cestode parasites other than T. ovis. 064-90
Species-specific, epitope of Chlamydia trachomatis and antibodies to it, protein sequence; application to diagnosis and as vaccine Mol. Diagnostics Eur 348 725; 3 January 1990 An epitope-specific peptide characterizing a common epitope of Chlamydia trachomatis (CT) having at least five amino acids is claimed. The peptide consists of at least four amino acids having a sequence position of I to XII of the epitope-specific natural sequence; Thr-Val-Phe-Asp-Val-Thr-Thr-Leu-Asn-ProThr-Ile. The four amino acids are in their sequence positions, but need not be contiguous to one another. The remaining amino acids are in their natural sequence or are immunogenic equivalents of the amino acids in their natural sequence. Also claimed is an anti-peptide antibody against a common epitope of CT, the antibody being produced by immunizing an animal against the peptide claimed. Also claimed is a nucleic acid probe for detecting CT-specific D N A or RNA sequences, consisting of <45 DNA or RNA bases and including a sequence encoding the specified protein sequence. The peptides characterize a species-specific epitope for serologically related serotypes of CT, and can be used in a vaccine for protection against CT infection or to detect CT in a test sample. 065-90
New recombinant D N A encoding HIV virus gag precursor protein without flanking sequences, expressed in insect, yeast and mammal cells; application to recombinant vaccine production Sk-Biol. Eur 345 242; 6 December 1989 New recombinant D N A (I) contains the a D N A sequence encoding the whole of the gag precursor protein of HIV virus, but lacking the natural 5' and 3' flanking sequences, operatively linked t o regulatory elements functional in eukaryotic cells. Also new are: (1) recombinant baculovirus and vaccinia virus vectors containing (I); (2) insect cells (Spodoptera frugiperda) infected with recombinant baculovirus; ( 3 ) m a m m a l cells (CHO, COS-7, NIH3T3, CV-1, mouse and rat myeloma, HAK, Vero, HeLa, W138, PRC-5) containing (I) or infected with (I)-containing vaccinia virus; and (4) recombinant yeast (Saccharomyces cerevisiae) cells containing (I). (I) induces expression of gag protein in the form of particles very similar in size and antigenic properties to those formed on the surface of infected human cells. Such particles lack viral functions required for maturation or replication, and are useful as anti-HIV virus vaccine components. 066-90
408 Vaccine, Vol. 8, August 1990
Vector encoding human cytomegalovirus glycoprotein; plasmid pSP65IT construction; HXLF1 and HXLF2 gene cloning and expression; recombinant vaccine construction; application in diagnosis and therapy Child. Hosp. St. Paul; Univ. lowa.-Res. Found. World. 8910 966; 16 November 1989 A recombinant expression vector comprises DNA derived from the human cytomegalovirus (HCMV) genome, where the DNA (HXLF1 and/or HXLF2 genes) encodes a portion of gp47-52 glycoprotein, which reacts with monoclonal antibody IVI10118. The immunogenic glycoproteins are of mol.wt 30 000 or 25 000, and may be used for recombinant vaccine construction. The expression vector is preferably a plasmid, containing the five homologous HCMV HXLF genes, or HXLF1 and/or HXLF2, and free of other HCMV genomic DNA. Nonglycosylated polypeptides encoded by HXLF1 have mol.wt 21 00025.000, and those encoded by HXLF1 and HXLF2 have mol.wt 20000. Plasmid pSP65IT contains the HXLF gene family cloned in plasmid SP6, and is deposited as plasmid IVI-10168. The plasmid contains an SP6 promoter and an ampicillinresistance selectable marker. Plasmid pSP64IT contains the genes in antisense DNA orientation. The recombinant proteins are useful in vaccines, monoclonal antibody and antigen-specific T-lymphocyte production for diagnosis or therapy of HCMV infection. 067-90
Production of Japanese-encephalitis virus envelope protein; expression of recombinant baculovirus vector e.g. nuclearpolyhedrosis virus in insect e.g. Spodoptera frugiperda cell culture; recombinant vaccine production and disease diagnosis Nippon-Zeon, Tokyo-Metrop. Inst. Neurosci. UK 2218 421; 15 November 1989 A new method for producing Japanese-encephalitis virus recombinant envelope protein comprises: (1) transforming insect cell cultures with baculovirus vectors, which contain at least a part of the Japanese-encephalitis virus eDNA; (2) culturing the transformed cells; and (3) recovering the recombinant envelope protein. The envelope protein eDNA is preferably associated with at least a part of the eDNA encoding the membrane protein of Japanese-encephalitis virus. Recombinant baculovirus containing the envelope, premembrane protein and/or membrane gene, and a vaccine preparation containing the envelope protein of Japanese-encephalitis virus are claimed. The new method produces large quantities of envelope protein, which can be used as a vaccine or for disease diagnosis. The vaccine dosage is sufficient to produce an antibody titre of 1-1(1oglo ). The baculovirus vector is preferably Autographa californica, Trichoplusia hi, Rachiplusia ou, Galleria mellonella or Bombyx mori nuclear-polyhedrosis virus. The preferred insect cells are Spodoptera frugiperda sf9 cells. 068-90
Glycoprotein of herpes zoster virus and its production; recombinant protein production in Saccharomyces cerevisiae for use in recombinant vaccine production K agaku-Oyob i-K essei-R yoho- K enkyusho Jpn 1252 279; 6 October 1989 A recombinant herpes zoster virus (HZV) glycoprotein is prepared by expressing the glycoprotein gpII gene in a eukaryotic cell. A vaccine against HZV infection, which is composed of the HZV glycoprotein, and an expression vector containing about 2.7kbp of HZV DNA are claimed. Transformants are cultured to prepare the recombinant glycoprotein. Recombinant HZV glycoprotein gpII has the same antigenicity as the virus-derived glycoprotein and may be used in HZV infection prophylaxis and as an assay reagent. The restriction endonuclease cleavage map of the gpII gene is given. The recombinant glycoprotein lacks a strongly hydrophobic region at the gpII-N-terminal domain. The host is preferably yeast (Saccharomyces cerevisiae). 069-90
Species-specific, epitope of Chlamydia trachomatis and antibodies to it, protein sequence; application to diagnosis and as vaccine Mol. Diagnostics Eur 348 725; 3 January 1990 An epitope-specific peptide characterizing a common epitope of Chlamydia trachomatis (CT) having at least five amino acids is claimed. The peptide consists of at least four amino acids having a sequence position of I to XII of the epitope-specific natural sequence; Thr-Val-Phe-Asp-Val-Thr-Thr-Leu-Asn-ProThr-Ile. The four amino acids are in their sequence positions, but need not be contiguous to one another. The remaining amino acids are in their natural sequence or are immunogenic equivalents of the amino acids in their natural sequence. Also claimed is an anti-peptide antibody against a common epitope of CT, the antibody being produced by immunizing an animal against the peptide claimed. Also claimed is a nucleic acid probe for detecting CT-specific D N A or RNA sequences, consisting of <45 DNA or RNA bases and including a sequence encoding the specified protein sequence. The peptides characterize a species-specific epitope for serologically related serotypes of CT, and can be used in a vaccine for protection against CT infection or to detect CT in a test sample. 065-90
New recombinant D N A encoding HIV virus gag precursor protein without flanking sequences, expressed in insect, yeast and mammal cells; application to recombinant vaccine production Sk-Biol. Eur 345 242; 6 December 1989 New recombinant D N A (I) contains the a D N A sequence encoding the whole of the gag precursor protein of HIV virus, but lacking the natural 5' and 3' flanking sequences, operatively linked t o regulatory elements functional in eukaryotic cells. Also new are: (1) recombinant baculovirus and vaccinia virus vectors containing (I); (2) insect cells (Spodoptera frugiperda) infected with recombinant baculovirus; ( 3 ) m a m m a l cells (CHO, COS-7, NIH3T3, CV-1, mouse and rat myeloma, HAK, Vero, HeLa, W138, PRC-5) containing (I) or infected with (I)-containing vaccinia virus; and (4) recombinant yeast (Saccharomyces cerevisiae) cells containing (I). (I) induces expression of gag protein in the form of particles very similar in size and antigenic properties to those formed on the surface of infected human cells. Such particles lack viral functions required for maturation or replication, and are useful as anti-HIV virus vaccine components. 066-90
408 Vaccine, Vol. 8, August 1990
Vector encoding human cytomegalovirus glycoprotein; plasmid pSP65IT construction; HXLF1 and HXLF2 gene cloning and expression; recombinant vaccine construction; application in diagnosis and therapy Child. Hosp. St. Paul; Univ. lowa.-Res. Found. World. 8910 966; 16 November 1989 A recombinant expression vector comprises DNA derived from the human cytomegalovirus (HCMV) genome, where the DNA (HXLF1 and/or HXLF2 genes) encodes a portion of gp47-52 glycoprotein, which reacts with monoclonal antibody IVI10118. The immunogenic glycoproteins are of mol.wt 30 000 or 25 000, and may be used for recombinant vaccine construction. The expression vector is preferably a plasmid, containing the five homologous HCMV HXLF genes, or HXLF1 and/or HXLF2, and free of other HCMV genomic DNA. Nonglycosylated polypeptides encoded by HXLF1 have mol.wt 21 00025.000, and those encoded by HXLF1 and HXLF2 have mol.wt 20000. Plasmid pSP65IT contains the HXLF gene family cloned in plasmid SP6, and is deposited as plasmid IVI-10168. The plasmid contains an SP6 promoter and an ampicillinresistance selectable marker. Plasmid pSP64IT contains the genes in antisense DNA orientation. The recombinant proteins are useful in vaccines, monoclonal antibody and antigen-specific T-lymphocyte production for diagnosis or therapy of HCMV infection. 067-90
Production of Japanese-encephalitis virus envelope protein; expression of recombinant baculovirus vector e.g. nuclearpolyhedrosis virus in insect e.g. Spodoptera frugiperda cell culture; recombinant vaccine production and disease diagnosis Nippon-Zeon, Tokyo-Metrop. Inst. Neurosci. UK 2218 421; 15 November 1989 A new method for producing Japanese-encephalitis virus recombinant envelope protein comprises: (1) transforming insect cell cultures with baculovirus vectors, which contain at least a part of the Japanese-encephalitis virus eDNA; (2) culturing the transformed cells; and (3) recovering the recombinant envelope protein. The envelope protein eDNA is preferably associated with at least a part of the eDNA encoding the membrane protein of Japanese-encephalitis virus. Recombinant baculovirus containing the envelope, premembrane protein and/or membrane gene, and a vaccine preparation containing the envelope protein of Japanese-encephalitis virus are claimed. The new method produces large quantities of envelope protein, which can be used as a vaccine or for disease diagnosis. The vaccine dosage is sufficient to produce an antibody titre of 1-1(1oglo ). The baculovirus vector is preferably Autographa californica, Trichoplusia hi, Rachiplusia ou, Galleria mellonella or Bombyx mori nuclear-polyhedrosis virus. The preferred insect cells are Spodoptera frugiperda sf9 cells. 068-90
Glycoprotein of herpes zoster virus and its production; recombinant protein production in Saccharomyces cerevisiae for use in recombinant vaccine production K agaku-Oyob i-K essei-R yoho- K enkyusho Jpn 1252 279; 6 October 1989 A recombinant herpes zoster virus (HZV) glycoprotein is prepared by expressing the glycoprotein gpII gene in a eukaryotic cell. A vaccine against HZV infection, which is composed of the HZV glycoprotein, and an expression vector containing about 2.7kbp of HZV DNA are claimed. Transformants are cultured to prepare the recombinant glycoprotein. Recombinant HZV glycoprotein gpII has the same antigenicity as the virus-derived glycoprotein and may be used in HZV infection prophylaxis and as an assay reagent. The restriction endonuclease cleavage map of the gpII gene is given. The recombinant glycoprotein lacks a strongly hydrophobic region at the gpII-N-terminal domain. The host is preferably yeast (Saccharomyces cerevisiae). 069-90