TY - JOUR
T1 - Cloning of differentially expressed genes in an HIV-1 resistant T cell clone by rapid subtraction hybridization, RaSH
AU - Simm, Malgorzata
AU - Su, Zao Zhong
AU - Huang, Eric Y.
AU - Chen, Yinming
AU - Jiang, Hongping
AU - Volsky, David J.
AU - Fisher, Paul B.
N1 - Funding Information:
We thank Dr M.J. Potash for fruitful discussions in the course of these studies. This research was supported in part by National Institutes of Health grants A144344, A143913 and NS31492, the Samuel Waxman Cancer Foundation and the Chernow Endowment. PBF is the Micahel and Stella Chernow Urological Cancer Research Scientist.
PY - 2001/5/16
Y1 - 2001/5/16
N2 - An HIV-1 resistant T cell clone R1c2 has been generated that carries mutant, latent HIV-1 in a minority of the cell population. Resistant cells express HIV-1 receptors CD4 and CXCR4 and display resistance to infection by wild type (wt) HIV-1 at the level of virus transcription. To begin to define the repertoire of genes modulated in R1c2 cells that correlate with and potentially control expression of the HIV-1 resistance phenotype we have employed a rapid subtraction hybridization (RaSH) technique. For this approach, cDNA libraries were prepared from double-stranded cDNAs that were enzymatically digested into small fragments, ligated to adapters, PCR amplified followed by incubation of tester and driver PCR fragments. The RaSH scheme resulted in the cloning of genes displaying differential expression between HIV-1 resistant (R1c2) and susceptible (SupT1) cells, including known genes and those not described in current DNA databases. Analysis of the pattern of expression of the differentially expressed genes documented eleven genes with enhanced (HR clones) and six genes with reduced (HS clones) expression in HIV-1 resistant versus HIV-1 susceptible T-cell clones.
AB - An HIV-1 resistant T cell clone R1c2 has been generated that carries mutant, latent HIV-1 in a minority of the cell population. Resistant cells express HIV-1 receptors CD4 and CXCR4 and display resistance to infection by wild type (wt) HIV-1 at the level of virus transcription. To begin to define the repertoire of genes modulated in R1c2 cells that correlate with and potentially control expression of the HIV-1 resistance phenotype we have employed a rapid subtraction hybridization (RaSH) technique. For this approach, cDNA libraries were prepared from double-stranded cDNAs that were enzymatically digested into small fragments, ligated to adapters, PCR amplified followed by incubation of tester and driver PCR fragments. The RaSH scheme resulted in the cloning of genes displaying differential expression between HIV-1 resistant (R1c2) and susceptible (SupT1) cells, including known genes and those not described in current DNA databases. Analysis of the pattern of expression of the differentially expressed genes documented eleven genes with enhanced (HR clones) and six genes with reduced (HS clones) expression in HIV-1 resistant versus HIV-1 susceptible T-cell clones.
KW - Differentially expressed sequence tags
KW - HIV-1 resistant and susceptible T cell clones
KW - Northern hybridization
KW - Reverse northern hybridization
UR - http://www.scopus.com/inward/record.url?scp=0035897668&partnerID=8YFLogxK
U2 - 10.1016/S0378-1119(01)00456-5
DO - 10.1016/S0378-1119(01)00456-5
M3 - Article
C2 - 11376941
AN - SCOPUS:0035897668
SN - 0378-1119
VL - 269
SP - 93
EP - 101
JO - Gene
JF - Gene
IS - 1-2
ER -