Comparison of the biological activities of human immunodeficiency virus 1 P24 and GP41 expressed in Spodoptera frugiperda cells by use of bac-to-bac system.
Recombinant transposing plasmids pFH24 and pFH41 were constructed by cloning the human immunodeficiency virus 1 (HIV-1) p24 and gp41 genes, respectively, into the transposing vector pFastBacHTa. Recombinant bacmids rBH24 and rBH41 were obtained by transposing pPolh/p24 and pPolh/gp41 expression cassettes from recombinant plasmids pFH24 and pFH41, respectively. Recombinant viruses rAcH24 and rAcH41 were generated by transfection of the Spodoptera frugiperda (Sf9) cells with the DNAs of plasmids rBH24 and rBH41, respectively. Analysis of the expressed p24 or gp41 proteins with an antiserum to HIV-1 (HIV-1 antiserum) by an enzyme-linked immunosorbent assay (ELISA) and dot blot assay showed high biological activity of these proteins; p24 was more active than gp41. Also a Western blot analysis showed stronger bands for p24 than for gp41. The high reactivities of p24 and gp41 with the HIV-1 antiserum suggest that these proteins could also be used as specific standard antigens in HIV-1 diagnostics.
Joshua
M N
MN
Institute of Virology, Wuhan University, Wuhan 430072, P.R. China.
Qi
Y
Y
Fu-Hua
Y
Y
Yong-Xiu
H
H
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors; D016655:HIV Core Protein p24 / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D015700:HIV Envelope Protein gp41 / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics* / Q000276:immunology; D006801:Humans; D007106:Immune Sera; D015151:Immunoblotting; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D018411:Spodoptera / Q000235:genetics*; D014162:Transfection
D016655:HIV Core Protein p24; D015700:HIV Envelope Protein gp41; D007106:Immune Sera; D011994:Recombinant Proteins