Screening and cloning for proteins transactivated by the PS1TP5 protein of hepatitis B virus: a suppression subtractive hybridization study.

ABSTRACT:

Aim

To clone and identify human genes transactivated by PS1TP5 by constructing a cDNA subtractive library with suppression subtractive hybridization (SSH) technique.

Methods

SSH and bioinformatics techniques were used for screening and cloning of the target genes transactivated by PS1TP5 protein. The mRNA was isolated from HepG2 cells transfected with pcDNA3.1(-)-myc-his(A)-PS1TP5 and pcDNA3.1(-)-myc-his(A) empty vector, respectively, and SSH technique was employed to analyze the differentially expressed DNA sequence between the two groups. After digestion with restriction enzyme Rsa I, small size cDNAs were obtained. Then tester cDNA was divided into two groups and ligated to the specific adaptor 1 and adaptor 2, respectively. The tester cDNA was hybridized with driver cDNA

SUBMITTER: Zhang JK 

PROVIDER: S-EPMC4146906 | biostudies-literature | 2007 Mar

REPOSITORIES: biostudies-literature