Expression of external glycoprotein gp105 and transmembrane glycoprotein gp36 of human immunodeficiency virus type 2 in insect cell by Bac-to-Bac system
2001
The recombinant transfer vector pFast Bac HTa-gp105 and pFast Bac HTb-gp36 were constructed by cloning HIV-2 gp105 gene and gp36 gene into the end of polyhedrin promoter of baculovirus transfer vector pFast Bac HTa and pFast Bac HTb respectively. HIV-2 external glycoprotein gp105 and transmembrane glycoprotein gp36 were expressed in insect cells Spodoptera frugiperda (Sf9) by transposing these two foreign genes to shuttle vector bacmid respectively using Bac-to-Bac system. The resulting products determined by SDS-PAGE showed that recombinant baculovirusescontaining gp105 gene expressed a 66 kDa glycoprotein, whereas those containing gp36 gene generated a 41 000u glycoprotein, the apparent molecular weight of which is equivalent to that of natural gp36. Western blot indicated that both gp105 and gp36 showed antigenic specificity. gp105 and gp36 has been expressed in insect cells Spodoptera frugiperda (Sf9) using baculovirus expression system.
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