Screening of the target genes transactivated by PS1TP1 protein with suppression subtractive hybridization technique
10.3760/cma.j.issn.1003-9279.2011.03.007
- VernacularTitle:应用抑制性消减杂交技术克隆PS1TP1的反式激活基因
- Author:
Hui-Huang HUANG
1
;
Dong JI
;
Si-Yu WANG
;
Wan-Zhen XU
;
Zhi-Yuan JIA
;
Ke LI
Author Information
1. 解放军第三0二医院
- Keywords:
Hepatitis B virus;
Protein precursors;
Protein,PS1TP1;
Transactivation(Genetics)
- From:
Chinese Journal of Experimental and Clinical Virology
2011;25(3):179-181
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a subtractive cDNA library of genes transactivated by PS1TP1 protein with suppression subtractive hybridization ( SSH) technique. Methods Suppression 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. Results The 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. Conclusion The 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 PS1TP1.
