J Bacteriol Virol.  2005 Jun;35(2):133-140.

Analysis on Quasispecies of HIV-1 env Gene by Single Clone per PCR

Affiliations
  • 1Center for AIDS Research, Department of Virology, National Institute of Health, Korea Center for Disease Control and Prevention, Seoul, Korea. jooshil@nih.go.kr
  • 2Biochemistry Lab., School of Life Science and Biotechnology, Korea Univ., Seoul, Korea.

Abstract

The human immunodeficiency viruses (HIV) exhibit tremendous genetic variability in their hosts. It is mainly due to two factors: the error-prone nature of the viral reverse transcriptase enzyme and the effects of environmental constraints such as antiviral therapy, cellular tropism, or HIV-specific immune responses. These quasispecies show fluctuation both in the overall divergence and diversity between individual sequences with different duration after primary infection. For better understand the viral quasispecies, we have performed the longitudinal genetic analysis of HIV-1 env gene V1-C5 region (1.2 kb) by two molecular cloning methods. Diversity indicated that 'single clone per PCR' value was higher than that of 'multiple clones per PCR' in subjects: 0.58-3.15 in subject 1 (P<0.05) and 0.28-2.25 in subject 2 (P<0.05). But divergence was similar in both molecular cloning methods. Phylogenetic analysis of longitudinal sequences at different sampling stages revealed the existence of different topologies individually. These data suggested that 'single clone per PCR' is more efficient than 'multiple clones per PCR' in genetic diversity analysis.

Keyword

HIV; env gene; Quasispecies; Molecular cloning

MeSH Terms

Clone Cells*
Cloning, Molecular
Genes, env*
Genetic Variation
HIV
HIV-1*
Polymerase Chain Reaction*
RNA-Directed DNA Polymerase
Tropism
RNA-Directed DNA Polymerase
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