| Literature DB >> 11112486 |
M O Salminen1, P K Ehrenberg, J R Mascola, D E Dayhoff, R Merling, B Blake, M Louder, S Hegerich, V R Polonis, D L Birx, M L Robb, F E McCutchan, N L Michael.
Abstract
We previously described the use of extended polymerase chain reaction (PCR) to amplify contiguous 9.2-kilobase (kb) single-long terminal repeat (LTR) proviral sequences from HIV-1 genetic subtypes A through G. We now extend these findings by describing a novel vector system to recover infectious molecular clones from long PCR amplicons. Directional ligation of 9.2-kb proviral amplicons into a recovery vector reconstitutes missing LTR sequences, providing candidate molecular clones for infectivity screening. We show that a previously characterized infectious molecular clone of HIV-1 retains its biological properties upon recovery with this strategy. Three additional infectious molecular clones generated, from primary isolates of subtype B (HIV-1(WR27)) and circulating recombinant form 01_AE (subtype E) (HIV-1(CM235)) by subtype-specific LTR reconstitution, displayed biological properties reflecting their cognate parental isolates. This represents the first report of infectious molecular clones from circulating recombinant form 01_AE (subtype E). Copyright 2000 Academic Press.Entities:
Mesh：
Substances：
Year: 2000 PMID： 11112486 DOI： 10.1006/viro.2000.0640
Source DB: PubMed Journal: Virology ISSN： 0042-6822 Impact factor: 3.616