Expression and Antigenicity of Human Immunodeficiency Virus Type-1 TransmembraneProtein GP41 in Insect Cells

摘要

The HIV-1 transmembrane protein, gp41, is processed together with the envelopeglycoprotein, gp120, from the same precursor, gp160, during the virus maturation. We used a baculovirus expression system to demonstrate that gp41 could be properly expressed without the preceding gp120 sequence. Two constructs with slight differences in the N-terminal region of gp41 were generated: one with a deletion of the first 7 hydrophobic residues of gp41, which have been suggested to be in a region important for membrane fusion and penetration, whereas the second with a complete sequence of gp41 except that a nonconserved leucine was substituted with a glutamine during DNA manipulation. Results from Western blotting with specific antisera confirm the gp41 identity. The sizes of gp41 were sensitive to tunicamycin treatment, indicating that N-linked glycosylation did occur. Further immunoblotting analyses with 90 different serum samples from HIV-1-infected individuals gave similar reaction patterns, suggesting that gp120 as well as the N-terminal region of gp41 are not critical for the expression and antigenecity of gp41. These eucaryotic constructs should provide valuable gp41 sources for detailed characterization of gp41 functions. Reprint, AIDS, HIV, Vaccines, Biotechnology, RAD I.