%0 Journal Article
%T Expression, purification, and bio-activity analysis of fusion protein HBx-EGFP-TLM<br>HBx-EGFP-TLM融合蛋白表达载体的构建、蛋白表达纯化及活性检测
%A Xiaoyan Shi
%A Yingying Zhang
%A Xiaowei Zhou
%A Jiansheng Lu
%A Zekun Guo
%A Peitang Huang
%A <br>史晓燕
%A 张莹莹
%A 周晓巍
%A 陆健昇
%A 郭泽坤
%A 黄培堂
%J 生物工程学报
%D 2011
%I 
%X Hepatitis B virus X protein (HBx) has various functions and plays a crucial role in the development of hepatocellular carcinoma (HCC). However, due to different transfection efficiency levels and experimental approaches, it is difficult to correlate the exact functions of HBx to HBV-associated HCC. In this study, we constructed two prokaryotic expression vectors, pGEX-HBx-EGFP-TLM and pGEX-EGFP-TLM, which expressed HBx-EGFP-TLM and EGFP-TLM fusion proteins respectively. Both vectors contained a coding sequence of TLM transduction motif derived from the PreS2-domain of Hepatitis B Virus surface antigens. In addition, EGFP was expressed as a reporter reflecting the transduction efficiency of TLM. The fusion protein HBx-EGFP-TLM or EGFP-TLM purified from Escherichia coli BL21(DE3) by ?KTATM Purifier system was incubated with AML12 and SMMC-7721 cells. Both Western blotting and laser confocal results indicated that the translocation motif TLM could lead HBx-EGFP and EGFP into the cytoplasm. Dual-Luciferase Reporter Assay revealed that the activity of mEZH2 promoter could be up-regulated by the recombinant HBx. In conclusion, we expressed a cell-permeable HBx, which could provide a new method to study the functions of HBx.
%K Hepatitis B virus
%K Hepatitis B virus X protein
%K prokaryotic expression
%K TLM translocation motif<br>乙型肝炎病毒，乙型肝炎病毒X蛋白，原核表达，转导肽TLM
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=F437B1BF01E7F486575CBE1B7D7ACAF6&yid=9377ED8094509821&vid=DB817633AA4F79B9&iid=9CF7A0430CBB2DFD&sid=40DE18199B7CA9BB&eid=9129323FE7AA9847&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=21