PMID- 10996639
OWN - NLM
STAT- MEDLINE
DCOM- 20001030
LR  - 20190831
IS  - 0166-0934 (Print)
IS  - 0166-0934 (Linking)
VI  - 89
IP  - 1-2
DP  - 2000 Sep
TI  - Distinct variation pattern in the env of macrophage-tropic simian 
      immunodeficiency virus in vivo demonstrated by denaturing gradient gel 
      electrophoresis.
PG  - 49-60
AB  - Genetic variations, occurring during the infection in the AIDS animal model using 
      a molecular clone virus, often consist of single nucleotide substitutions 
      distributed sparsely in the viral genome. A highly sensitive method was developed 
      to detect such mutations, consisting of denaturing gradient gel electrophoresis 
      (DGGE) and direct sequencing. The genetic variation in the env of a macrophage 
      tropic simian immunodeficiency virus (SIV) in a macaque infected chronically was 
      examined by this technique and compared with that of a T cell tropic SIV in an 
      animal infected chronically. A whole env sequence was amplified by 11 sets of PCR 
      for DNA ranging from 300 to 420 bp. The amplified DNA was subjected to mutational 
      screening by DGGE and to subsequent direct sequencing. Imaging analysis of 
      separated DNA bands by DGGE provided information on the proportion of each 
      variant DNA. The validity of this technique was confirmed by sequencing of 
      variant DNAs cloned by limiting dilutions. Thus, this technique is suitable for 
      analysis of genetic variations in the AIDS animal model using a molecular clone 
      virus.
FAU - Dhar, S K
AU  - Dhar SK
AD  - Tsukuba Primate Center for Medical Sciences, National Institute for Infectious 
      Diseases, 1 Hachimandai, 305-0843, Tsukuba, Japan.
FAU - Tadakuma, K
AU  - Tadakuma K
FAU - Mori, K
AU  - Mori K
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - J Virol Methods
JT  - Journal of virological methods
JID - 8005839
RN  - 0 (DNA, Viral)
RN  - 0 (Gene Products, env)
SB  - IM
MH  - Animals
MH  - Chronic Disease
MH  - DNA Mutational Analysis/methods
MH  - DNA, Viral/*analysis
MH  - Electrophoresis, Polyacrylamide Gel/methods
MH  - Female
MH  - Gene Products, env/*genetics
MH  - Macaca
MH  - Macrophages/*virology
MH  - Male
MH  - Polymerase Chain Reaction
MH  - Polymorphism, Single Nucleotide
MH  - Simian Acquired Immunodeficiency Syndrome/*virology
MH  - Simian Immunodeficiency Virus/*genetics
MH  - T-Lymphocytes/*virology
MH  - Tropism
EDAT- 2000/09/21 11:00
MHDA- 2001/02/28 10:01
CRDT- 2000/09/21 11:00
PHST- 2000/09/21 11:00 [pubmed]
PHST- 2001/02/28 10:01 [medline]
PHST- 2000/09/21 11:00 [entrez]
AID - S0166093400002019 [pii]
AID - 10.1016/s0166-0934(00)00201-9 [doi]
PST - ppublish
SO  - J Virol Methods. 2000 Sep;89(1-2):49-60. doi: 10.1016/s0166-0934(00)00201-9.