隱源性肝炎相關(guān)新基因CHBP2的原核表達及蛋白純化
隱源性肝炎;CHBP2;原核表達;蛋白純化,李錕,葉進,肖凡,李國力,洪源,魏紅山,通訊作者:,ExpressionandpurificationofnewgeneCHBP2associatedwithcryptogenichepatitisi
 |
| 第1頁 |
參見附件(612KB，5頁)。
李錕, 葉進, 華中科技大學(xué)同濟醫(yī)學(xué)院附屬協(xié)和醫(yī)院消化內(nèi)科 湖北省武漢市 430022
肖凡, 李國力, 洪源, 魏紅山, 北京地壇醫(yī)院傳染病研究室 北京市 100011
李錕, 華中科技大學(xué)同濟醫(yī)學(xué)院2004級碩士研究生, 主治醫(yī)師, 主要從事肝病的分子生物學(xué)及臨床研究.
通訊作者: 葉進, 430022, 湖北省武漢市, 華中科技大學(xué)同濟醫(yī)學(xué)院附屬協(xié)和醫(yī)院消化內(nèi)科. yejin8688@sina.com
電話: 027-85726381
收稿日期: 2007-03-04 修回日期: 2007-07-18
Expression and purification of new gene CHBP2 associated with cryptogenic hepatitis in prokaryote cells
Kun Li, Jin Ye, Fan Xiao, Guo-Li Li, Yuan Hong, Hong-Shan Wei
Kun Li, Jin Ye, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China
Fan Xiao, Guo-Li Li, Yuan Hong, Hong-Shang Wei, Institute of Infectious Diseases, Ditan Hospital, Beijing 100011, China
Correspondence to: Dr. Jin Ye, Department of Digestive Disease, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China. yejin8688@sina.com
Received: 2007-03-04 Revised: 2007-07-18
Abstract
AIM: To construct a prokaryotic cell expression vector for a new gene, CHBP2, associated with cryptogenic hepatitis, and to abundantly express and purify CHBP2 protein.
METHODS: The open reading frame (ORF) of CHBP2 was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR), in which an mRNA template was extracted from Huh7 cells and ligated into the pGEM-T cloning vector. After sequencing, the correct DNA fragment was inserted into inducible Escherichia coli BL21. After the CHBP2 protein was induced with Isopropyl-beta-d-thiogalactopyranoside (IPTG), it was analyzed by SDS-PAGE and western blotting. Expressed bacteria were atomized by ultrasound and analyzed with SDS-PAGE. The expressed product was purified and renatured by Ni+ affinity column chromatography.
RESULTS: The prokaryotic cell expression vector pET-32a(+)-CHBP2 was successfully constructed and CHBP2 protein was abundantly expressed ......
您現(xiàn)在查看是摘要介紹頁，詳見PDF附件(612KB，5頁)。