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© Marie-Christine Prévost, Nathalie Sol-Foulon, Olivier Schwartz, Jean-Marc Panaud
AIDS virus particles at the surface of a lymphocyte.
Publication : The Journal of general virology

Recovery of APOBEC3-edited human immunodeficiency virus G->A hypermutants by differential DNA denaturation PCR

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in The Journal of general virology - 01 Jan 2005

Suspène R, Henry M, Guillot S, Wain-Hobson S, Vartanian JP

Link to Pubmed [PMID] – 15604439

J. Gen. Virol. 2005 Jan;86(Pt 1):125-9

Virus genomes from the same family may exhibit a wide range in their DNA GC content, whereas viral hypermutants differ substantially in GC content from their parental genomes. As AT-rich DNA melts at lower temperatures than GC-rich DNA, use of a lower denaturation temperature during PCR should allow differential amplification of AT-rich genomes or variants within a quasispecies. The latter situation has been explored explicitly in a two-step process by using a series of well-defined viral sequences differing in their AT content. Firstly, the lowest denaturation temperature (T(p)) that allowed amplification of the parental sequence was determined. Secondly, differential amplification of AT-rich viral variants was obtained by using a denaturation temperature 1-3 degrees C lower than T(p). Application of this sensitive method to two different viruses allowed us to identify human immunodeficiency virus type 1 G–>A hypermutants in a situation where none were expected and to amplify AT-rich variants selectively within a spectrum of poliovirus mutants.

http://www.ncbi.nlm.nih.gov/pubmed/15604439