The invention discloses a potential preparation method for a highly-efficient recombinant HIV-1 CRF07-BC gp140
immunogen. The
immunogen is designed based on the structure of HIV-1 envelope
protein crystals which have been published internationally and obtained in our lab. The specific method uses an overlap extension PCR technology to obtain gp140
gene segments, and comprises the steps that target genes are cloned into an eukaryotic
expression vector pMT, endotoxin is removed through extraction of a large amount of plasmids, the gp140
gene segments and resistance screening plasmids pCoBlast are co-transfected into
drosophila melanogaster Schneider2 (S2) cells together, blasticidin (Blasticidin S) is used for positive clone screening, S2
cell lines for stably and efficiently secreting and expressing gp140 are screened, and after enlarged cultivation and through two steps of purification of
nickel column
affinity chromatography and gel
filtration chromatography, the gp140 with high purity can be obtained. A series of biochemical and biophysical technologies indicate that the gp140 is uniform in polymeric states, and high in antigenic reactivity, and is quite fittingly used as the
immunogen for the research and the development of AIDS
subunit vaccines or multivalent
combined vaccines.