Expression of the extracellular domain of the human immunodeficiency virus type 1 envelope protein and its fusion with beta-galactosidase in Saccharomyces cerevisiae.
Two envelope glycoprotein gene fragments were cloned from the proviral genome of the HXB2 isolate of human immunodeficiency virus (HIV). For the production of the two domains of the envelope gene product these cloned gene fragments were inserted into an Escherichia coli-yeast inducible shuttle vector fused to the galactokinase (GAL1) promoter. Cell extracts from strains of Saccharomyces cerevisiae harboring these two vectors (pYENV1 and pYENV2) were found to contain a specific protein with a size of 50 kDa when induced by galactose, while the protein could not be detected in extracts from control cells containing only the E. coli-yeast vector in the presence of galactose. Furthermore, another expression plasmid coding for fusion proteins from the majority of the external envelope glycoprotein (gp120) moiety and a large part of the beta-galactosidase was constructed. Antibodies from HIV type 1-positive sera could react with recombinant fusion polypeptides. Transformants could produce this fusion protein to a level of about 1.6% of the total protein content, as deduced from beta-galactosidase activity.
State Key Laboratory of Microbial Technology, College of Life Science, Shandong University, Jinan, China. weifengl@life.sdu.edu.cn
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Liu
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Gao
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Wang
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005690:Galactose / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D015686:Gene Products, env / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D015752:Genes, env; D005822:Genetic Vectors; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120 / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D015497:HIV-1 / Q000235:genetics* / Q000276:immunology; D006801:Humans; D010446:Peptide Fragments / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D011533:Proviruses / Q000235:genetics / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D012441:Saccharomyces cerevisiae / Q000235:genetics; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D015686:Gene Products, env; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase; D005690:Galactose