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Science 5 April 1985:
Vol. 228. no. 4695, pp. 93 - 96
DOI: 10.1126/science.2983429
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Articles

Science, Vol 228, Issue 4695, 93-96
Copyright © 1985 by American Association for the Advancement of Science

articles

Expression in Escherichia coli of open reading frame gene segments of HTLV-III

NT Chang, PK Chanda, AD Barone, S McKinney, DP Rhodes, SH Tam, CW Shearman, J Huang, TW Chang, RC Gallo, and al. et

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.


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