TY - JOUR
T1 - Construction and biological characterization of infectious molecular clones of HIV-1 subtypes B and E (CRF01-AE) generated by the polymerase chain reaction
AU - Salminen, Mika O.
AU - Ehrenberg, Philip K.
AU - Mascola, John R.
AU - Dayhoff, Deborah E.
AU - Merling, Randall
AU - Blake, Billy
AU - Louder, Mark
AU - Hegerich, Susan
AU - Polonis, Victoria R.
AU - Birx, Deborah L.
AU - Robb, Merlin L.
AU - McCutchan, Francine E.
AU - Michael, Nelson L.
N1 - Funding Information:
We thank Eric Sanders-Buell and Carol Wang for technical assistance, and Jean Carr, Richard Carroll, Jim Riley, and Dan St. Louis for helpful discussions. The views and opinions expressed herein are those of the authors and do not purport to reflect the official policy or position of the U.S. Army or the Department of Defense. This work was supported, in part, by Cooperative Agreement No. DAMD17-93-V-3004, between the U.S. Army Medical Research and Materiel Command and the Henry M. Jackson Foundation for the Advancement of Military Medicine. M. Salminen was partly supported by Grant 19191 from the Finnish Academy of Science.
PY - 2000/12/5
Y1 - 2000/12/5
N2 - 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(WR237)) 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). (C) 2000 Academic Press.
AB - 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(WR237)) 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). (C) 2000 Academic Press.
UR - http://www.scopus.com/inward/record.url?scp=0034610249&partnerID=8YFLogxK
U2 - 10.1006/viro.2000.0640
DO - 10.1006/viro.2000.0640
M3 - Article
C2 - 11112486
AN - SCOPUS:0034610249
SN - 0042-6822
VL - 278
SP - 103
EP - 110
JO - Virology
JF - Virology
IS - 1
ER -