[High-level expression of human immunodeficiency virus type 1 gp120 protein in E. coli].
In order to improve protein expression and to produce good and cheap diagnostic antigen, the gene fragment (560bp) in N-terminal of gp120 of HIV-1 LAV strain was amplified by PCR. After digested by EcoR I and Sal I, the fragment was cloned into a high-level expression vector pET28a. The recombinant plasmid pET/120 transfecting BL21 (DE3) produced the protein with high-level expression in the host cell BL21(DE3), which was further proved having good antigenicity and high specificity by indirect ELISA and Western-blot assay. The protein expressed was about 50% of the total bacterial protein by SDS-PAGE electrophoresis test. It was highly expressed in the prokaryotic expression system.
Bi
L
L
Wuhan Institute of Biological Products, Ministry of Health, Wuhan 430060.
Lei
X
X
Zhu
J
J
D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005814:Genes, Viral; D005822:Genetic Vectors; D015699:HIV Envelope Protein gp120 / Q000096:biosynthesis* / Q000235:genetics; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011387:Prokaryotic Cells; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
D004274:DNA, Recombinant; D015699:HIV Envelope Protein gp120; D011994:Recombinant Proteins