Bacterial expression and characterization of nine polypeptides encoded by segments of the envelope gene of human immunodeficiency virus.
Nine envelope (Env) polypeptides, encoding different regions of HIV gp120 and gp41 Env proteins, and accounting for approx. 96% of the entire Env precursor glycoprotein complex (gp160) were expressed in Escherichia coli at levels ranging from approx. 2 to 20% of total cellular protein. The recombinant polypeptides were produced either as hybrid products fused to the cII gene fragment of the lambda vector or in an unfused form without interfering cII products. Partially purified protein fractions of each polypeptide were characterized serologically by Western-blot analysis against a panel of well characterized human immunodeficiency virus (HIV)-positive human reference sera. Most of the Env polypeptides were highly immunoreactive with anti-gp120/gp41 antibodies present in the sera of patients with acquired immunodeficiency syndrome (AIDS) and AIDS-related diseases, but the patterns of reactivity were different. These results demonstrate that some of the antigenic determinants residing on the viral gp160 complex are retained on the surfaces of the recombinant Env polypeptides, and suggest that these sites are differentially immunogenic. These results are therefore interpreted in the context of an ongoing process towards using bacterially expressed HIV Env polypeptides to help define biological and structural epitopes to aid in the development of more sensitive diagnostic and therapeutic reagents in the fight against AIDS.
Samuel
K P
KP
Laboratory of Molecular Oncology, NCI-Frederick Cancer Research Facility, MD 21701.
Seth
A
A
Zweig
M
M
Showalter
S D
SD
Papas
T S
TS
D003001:Cloning, Molecular / Q000379:methods; D004262:DNA Restriction Enzymes; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D005814:Genes, Viral; D005822:Genetic Vectors; D006678:HIV / Q000235:genetics*; D010957:Plasmids; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000302:isolation & purification
D014759:Viral Envelope Proteins; D004262:DNA Restriction Enzymes