title: Expression in Escherichia coli of open reading frame gene segments of HTLV-III.
abstract: 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.
authors.forename: N T
authors.initials: NT
authors.lastname: Chang
authors.forename: P K
authors.initials: PK
authors.lastname: Chanda
authors.forename: A D
authors.initials: AD
authors.lastname: Barone
authors.forename: S
authors.initials: S
authors.lastname: McKinney
authors.forename: D P
authors.initials: DP
authors.lastname: Rhodes
authors.forename: S H
authors.initials: SH
authors.lastname: Tam
authors.forename: C W
authors.initials: CW
authors.lastname: Shearman
authors.forename: J
authors.initials: J
authors.lastname: Huang
authors.forename: T W
authors.initials: TW
authors.lastname: Chang
authors.forename: R C
authors.initials: RC
authors.lastname: Gallo
mesh_terms: 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
chemical_list: D000914:Antibodies, Viral; D004274:DNA, Recombinant; D004279:DNA, Viral; D014764:Viral Proteins; D001616:beta-Galactosidase