[Cloning and expression of the envelope glycoprotein gD gene of pseudorabies virus EA strain].
The envelope glycoprotein gD gene of pseudorabies virus Ea strain was cloned via PCR technique. Sequence analysis displayed 98% nucleotide sequence homology and 97% deduced amino acid sequence homology between our cloned gD gene and PRV Rice strain gD gene. The recombinant transfer plasmid pSX35A-gD was obtained by inserting D gene into the baculovirus transfer vector pSX35A with whole-phase promoter cassette, then transfected insect cell Hi5 with linearized AcMNPV-OCC- virus DNA, and formed recombinant baculoviruses AcMNPV-OCC(+)-gD by homologous recombination in insect cell. Recombinant baculoviruses infected insect cell Hi5 after being purified by plaque assay. Both SDS-PAGE and Western-blotting showed glycoprotein gD with a molecular weight of about 47 kD was expressed specifically, product was about 6.2% of total cellular protein, and expressed gD was of immunogenicity.
Chen
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State Key Lab for Biocontrol, Zhongshan University, Guangzhou 510275, China.
Yang
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Hong
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Chen
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H
Chen
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Long
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Wang
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X
D000818:Animals; D016367:Baculoviridae / Q000378:metabolism; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics; D005814:Genes, Viral; D011558:Herpesvirus 1, Suid / Q000737:chemistry / Q000145:classification / Q000235:genetics*; D009036:Moths / Q000166:cytology / Q000378:metabolism; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics*; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics
D004279:DNA, Viral; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C107752:glycoprotein D, pseudorabies virus