Human immunodeficiency virus type-1 p24 sequence from an Indian strain: expression in Escherichia coli and implications in diagnostics.
A 637-bp fragment, corresponding to the p24 human immunodeficiency virus (HIV) core protein from the gag ORF, was PCR amplified from DNA isolated from peripheral blood mononuclear lymphocytes (PBML) of an asymptomatic HIV-1 seropositive human subject from Bombay and cloned into PCRScript SK(+). The nucleotide sequence revealed highest homology (98.6%) with the consensus sequence of the HIV-1 B subtype. The 637-bp KpnI-HindIII fragment was cloned downstream from a His6 tag in the pQE30 vector under the control of phage T5 promoter leading to production of a 6XHis-p24 fusion protein in Escherichia coli. It showed an approx. 24-kDa band by SDS-PAGE. The recombinant p24 reacted with serum samples from HIV-infected subjects when tested by Western blot and ELISA.
Gupta
S K
SK
Gamete Antigen Laboratory, National Institute of Immunology, New Delhi, India. GA@nii.ernet.in
Sengupta
J
J
Bisht
R
R
Bhatnagar
A
A
Kaul
R
R
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004279:DNA, Viral; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D016655:HIV Core Protein p24 / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D007194:India; D008969:Molecular Sequence Data; D013045:Species Specificity
D004279:DNA, Viral; D016655:HIV Core Protein p24