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Expression in Escherichia coli of open reading frame gene segments of HTLV-III | | N T Chang
; P K Chanda
; A D Barone
; S McKinney
; D P Rhodes
; S H Tam
; C W Shearman
; J Huang
; T W Chang
; R C Gallo
; | | Date: |
5 Apr 1985 | | Journal: | Science, 228 (4695), 93-6 | | 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. | | Source: | PubMed, pmid2983429 | | Services: | Forum | Review | Favorites | |
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