Recombinant polypeptide from the endonuclease region of the acquired immune deficiency syndrome retrovirus polymerase (pol) gene detects serum antibodies in most infected individuals.
Sera from the majority of individuals that were positive in an enzyme-linked immunosorbent assay (ELISA) retrovirus (ARV), an isolate of the for antibodies to acquired immune deficiency syndrome (AIDS)-associated retrovirus (ARV), an isolate of the retrovirus identified as the etiologic agent of AIDS, were found to react with a 31,000-dalton protein (p31) in virus Western blot assays. To determine if this 31,000-dalton immunoreactive species originated from the putative endonuclease region of the polymerase (pol) gene of ARV, we cloned this portion of pol into bacterial expression vectors for direct expression and for expression as a fusion protein with human superoxide dismutase. Transformants from both constructions expressed immunoreactive protein detected in immunoblots with an AIDS patient's serum. Extracts from transformants expressing these sequences competed with the binding of antibodies from AIDS patients' sera to the 31,000-dalton protein in virus immunoblots, confirming that viral p31 originated from the endonuclease domain of the ARV polymerase gene. The superoxide dismutase-p31 fusion protein was purified, and an ELISA for detecting antibodies to p31 was developed. The majority (95%) of serum samples obtained from individuals seropositive in the virus ELISA were also positive in the p31 antibody ELISA.
Steimer
K S
KS
Higgins
K W
KW
Powers
M A
MA
Stephans
J C
JC
Gyenes
A
A
George-Nascimento
C
C
Luciw
P A
PA
Barr
P J
PJ
Hallewell
R A
RA
Sanchez-Pescador
R
R
D000163:Acquired Immunodeficiency Syndrome / Q000175:diagnosis / Q000276:immunology* / Q000382:microbiology; D000914:Antibodies, Viral / Q000276:immunology; D001667:Binding, Competitive; D003001:Cloning, Molecular; D017977:Deltaretrovirus / Q000235:genetics / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D005814:Genes, Viral; D006801:Humans; D007163:Immunosorbent Techniques; D008970:Molecular Weight; D012194:RNA-Directed DNA Polymerase / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology*; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology*
D000914:Antibodies, Viral; D011994:Recombinant Proteins; D014764:Viral Proteins; D012194:RNA-Directed DNA Polymerase