Construction and Expression of Hepatitis B Virus X Gene Expression Vector in Eucaryotic Cells
10.3321/j.issn:1000-467X.2001.05.012
- VernacularTitle:乙肝病毒 X基因真核表达载体的构建及表达
- Author:
Xiao-Zhong WANG
1
;
Zhi-Xin CHEN
;
Yue-Hong HUANG
;
Xiao-Chun CHEN
;
Qi-Min TAO
Author Information
1. Union Hospital Fujian Medical University
- From:Chinese Journal of Cancer
2001;20(5):499-501
- CountryChina
- Language:Chinese
-
Abstract:
Objective： This study was designed to construct the eukaryotic expression vector of HBV X gene for exploring the role of X gene in the carcinogenesis of hepatocellular carcinoma. Methods： X gene containing EcoR Ⅰ and Pst Ⅰ endoenzyme sites was obtained using PCR; Double enzyme digestion was conducted for vector PAS2-1 and PCR product of X gene; Both fragments were connected using ligase and transferred to Eco-JM105; Reconstitute plasmid sequence was examined by auto-sequencing assay and was named as PAS2-1X; Reconstituted plasmid was transformated into the yeast cell AH109 by Liac-mediated transformation; The X protein expressed in the yeast cell was confirmed by Western blot analysis. Results： Reconstituted plasmid PAS2-1X included the anticipated fragment of X gene was proved by auto-sequencing assay. Western blot analysis showed that reconstitute plasmid PAS2-1X can express the X protein in the yeast cell. Conclusion： Reconstitution of PAS2-1X vector laid a foundation for better understanding of the mechanism of HBV X gene and X-protein in carcinogenesis of hepatocellular carcinoma, and was helpful for finding the X-related protein that was supposed to react to X-protein in vivo by yeast two-hybrid system.
