Expression in Escherichia coli of open reading frame gene segments of HTLV-III.
Human T-cell lymphotropic virus type III (HTLV-III), the causative agent of the acquired immune deficiency syndrome (AIDS), was recently isolated and its genomic structure analyzed by DNA cloning methods. In the studies reported here a combined cloning and expression system was used to identify HTLV-III encoded peptides that react immunologically with antibodies in sera from AIDS patients. Cloned HTLV-III DNA was sheared into approximately 500-base-pair fragments and inserted into an "open reading frame" expression vector, pMR100. The inserted DNA was expressed in Escherichia coli transformants as a polypeptide fused to the lambda CI protein at its amino terminus and to beta-galactosidase at its carboxyl terminus. Sera from AIDS patients containing antibodies to HTLV-III were then used to screen for immunoreactive fusion proteins. Twenty clones, each specifying a fusion protein strongly reactive with AIDS serum, were identified. DNA sequence analysis indicated that the HTLV-III fragments were derived from the open reading frame DNA segments corresponding to the gag and pol gene coding regions and also the large open reading frame region (env-lor) located near the 3' end of the viral genome.
Chang
N T
NT
Chanda
P K
PK
Barone
A D
AD
McKinney
S
S
Rhodes
D P
DP
Tam
S H
SH
Shearman
C W
CW
Huang
J
J
Chang
T W
TW
Gallo
R C
RC
D000163:Acquired Immunodeficiency Syndrome / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000378:metabolism; D004279:DNA, Viral / Q000235:genetics; D017977:Deltaretrovirus / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005814:Genes, Viral; D005822:Genetic Vectors; D006801:Humans; D014764:Viral Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D001616:beta-Galactosidase / Q000378:metabolism
D000914:Antibodies, Viral; D004274:DNA, Recombinant; D004279:DNA, Viral; D014764:Viral Proteins; D001616:beta-Galactosidase