Screening of the target genes transactivated by PS1TP1 protein with suppression subtractive hybridization technique.
- Author:
Hui-Huang HUANG
1
;
Dong JI
;
Si-Yu WANG
;
Wan-Zhen XU
;
Zhi-Yuan JIA
;
Ke LI
Author Information
- Publication Type:Journal Article
- MeSH: Hepatitis B; genetics; Humans; Nucleic Acid Hybridization; methods; Transcriptional Activation; genetics; Viral Proteins; genetics
- From: Chinese Journal of Experimental and Clinical Virology 2011;25(3):179-181
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct a subtractive cDNA library of genes transactivated by PS1TP1 protein with suppression subtractive hybridization (SSH) technique.
METHODSSuppression subtractive hybridization technique and bioinformatics technique were used, the mRNA from HepG2 cells transfected with pcDNA3.1 (-) -PS1TP1 and pcDNA3. 1 (-) empty vector was isolated, respectively; cDNA underwent two times of nested PCR, amplified cDNA fragments were subcloned into pGEM-Teasy vectors to set up the subtractive library.
RESULTSThe subtractive library of genes transactivated by PS1TP1 was constructed successfully. Sequence analysis was performed in 43 clones randomly, and the full length sequences were obtained with bioinformatics method and searched for homologous DNA sequence from GenBank, altogether 12 coding sequences were gotten, which consisted of 10 known and 2 unknown ones.
CONCLUSIONThe obtained sequences may be target genes transactivated by PS1TP1 protein among which some genes coding proteins involved in cell cycle regulation, metabolism, immunity and cell apoptosis. This finding brought some clues for studying the biological functions of PS1T1.
