LIN Shuª²Mei1, ZHANG Shuª²Lin1, CHENG Jun2, LIU Min1£¬ GUO Jiang2£¬ ZHANG Liª²Ying2£¬ YANG Yuan1

    1Department of Infectious Diseases, First Affiliated Hospital, Medical College£¬ Xi¬ðan Jiaotong University, Xi¬ðan 710061, China, 2Institute of Infectious Diseases, Beijing Ditan Hospital, Beijing 100011, China

    ¡¾Abstract¡¿ AIM£º To clone and identify human genes transactivated by human gene C1 encoding protein C1 interacting with hepatitis B virus (HBV) core antigen by constructing a cDNA subtractive library with suppression subtractive hybridization (SSH) technique. METHODS£º SSH and bioinformatics techniques were used for screening and cloning of the target genes transactivated by C1 protein. The mRNA was isolated from HepG2 cells transfected with pcDNA3.1(-)ª²C1 and pcDNA3.1(-) empty vector, respectively, SSH method was employed to analyze the differentially expressed cDNA sequence between the 2 groups. After restriction enzyme Rsa I digestion, small sizes cDNAs were obtained. Then tester cDNA was divided into 2 groups and ligated to the specific adaptor 1 and adaptor 2, respectively. After tester cDNA was hybridized with driver cDNA twice and underwent polymerase chain reaction (PCR) twice and then was subcloned into pGEMª²Teasy plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E. coli strain DH5¦Á. The cDNA was sequenced and analyzed in GenBank with BLAST search after PCR. RESULTS£º The subtractive library of genes transactivated by C1 was constructed successfully. Colony PCR of 40 positive clones showed that these clones contained 200-1000 bp inserts. Sequence analysis was performed in 30 clones ......