Construction and biological characterization of infectious molecular clones of HIV-1 subtypes B and E (CRF01-AE) generated by the polymerase chain reaction

Mika O. Salminen; Philip K. Ehrenberg; John R. Mascola; Deborah E. Dayhoff; Randall Merling; Billy Blake; Mark Louder; Susan Hegerich; Victoria R. Polonis; Deborah L. Birx; Merlin Robb; Francine E. McCutchan; Nelson L. Michael

doi:10.1006/viro.2000.0640

Construction and biological characterization of infectious molecular clones of HIV-1 subtypes B and E (CRF01-AE) generated by the polymerase chain reaction

Mika O. Salminen
, Philip K. Ehrenberg
, John R. Mascola
, Deborah E. Dayhoff
, Randall Merling
, Billy Blake
, Mark Louder
, Susan Hegerich
, Victoria R. Polonis
, Deborah L. Birx
, Merlin Robb
, Francine E. McCutchan
, Nelson L. Michael^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

23 Scopus citations

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(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.

Original language	English
Pages (from-to)	103-110
Number of pages	8
Journal	Virology
Volume	278
Issue number	1
DOIs	https://doi.org/10.1006/viro.2000.0640
State	Published - 5 Dec 2000

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10.1006/viro.2000.0640

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Salminen, M. O., Ehrenberg, P. K., Mascola, J. R., Dayhoff, D. E., Merling, R., Blake, B., Louder, M., Hegerich, S., Polonis, V. R., Birx, D. L., Robb, M., McCutchan, F. E., & Michael, N. L. (2000). Construction and biological characterization of infectious molecular clones of HIV-1 subtypes B and E (CRF01-AE) generated by the polymerase chain reaction. Virology, 278(1), 103-110. https://doi.org/10.1006/viro.2000.0640

@article{086c05b40e1341978a7ed12162b73245,

title = "Construction and biological characterization of infectious molecular clones of HIV-1 subtypes B and E (CRF01-AE) generated by the polymerase chain reaction",

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(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.",

author = "Salminen, \{Mika O.\} and Ehrenberg, \{Philip K.\} and Mascola, \{John R.\} and Dayhoff, \{Deborah E.\} and Randall Merling and Billy Blake and Mark Louder and Susan Hegerich and Polonis, \{Victoria R.\} and Birx, \{Deborah L.\} and Merlin Robb and McCutchan, \{Francine E.\} and Michael, \{Nelson L.\}",

note = "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.",

year = "2000",

month = dec,

day = "5",

doi = "10.1006/viro.2000.0640",

language = "English",

volume = "278",

pages = "103--110",

journal = "Virology",

issn = "0042-6822",

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Salminen, MO, Ehrenberg, PK, Mascola, JR, Dayhoff, DE, Merling, R, Blake, B, Louder, M, Hegerich, S, Polonis, VR, Birx, DL, Robb, M, McCutchan, FE & Michael, NL 2000, 'Construction and biological characterization of infectious molecular clones of HIV-1 subtypes B and E (CRF01-AE) generated by the polymerase chain reaction', Virology, vol. 278, no. 1, pp. 103-110. https://doi.org/10.1006/viro.2000.0640

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

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 - https://www.scopus.com/pages/publications/0034610249

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 -

Construction and biological characterization of infectious molecular clones of HIV-1 subtypes B and E (CRF01-AE) generated by the polymerase chain reaction

Abstract

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