Expression and antigenecity of human immunodeficiency virus type-1 transmembrane protein gp41 in insect cells.
The HIV-1 transmembrane protein, gp41, is processed together with the envelope glycoprotein, 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.
Yu
S L
SL
Graduate Institute of Microbiology & Immunology, Natl. Yang-Ming Medical College, Taipei, Taiwan.
Chou
M J
MJ
Tam
M F
MF
Lee
T H
TH
Syu
W J
WJ
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D000939:Epitopes / Q000032:analysis; D015870:Gene Expression / Q000187:drug effects; D005822:Genetic Vectors; D015699:HIV Envelope Protein gp120 / Q000235:genetics; D015700:HIV Envelope Protein gp41 / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D015497:HIV-1 / Q000235:genetics*; D007313:Insecta; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D015183:Restriction Mapping; D014162:Transfection; D014415:Tunicamycin / Q000494:pharmacology
D000939:Epitopes; D015699:HIV Envelope Protein gp120; D015700:HIV Envelope Protein gp41; D009838:Oligodeoxyribonucleotides; D014415:Tunicamycin