枯草芽孢杆菌的ade基因在大肠杆菌中的MBP融合表达及活性鉴定

枯草芽孢杆菌的ade基因在大肠杆菌中的MBP融合表达及活性鉴定

付玉芹, 李敏, 岳晓婧, 崔银秋, 周慧

（吉林大学生命科学学院大分子实验室，长春 130021）

收稿日期:2005-08-26 修回日期:1900-01-01 出版日期:2006-03-26 发布日期:2006-03-26
通讯作者: 崔银秋

Fusion Expression and Characterization of the ade Gene from Bacillus Subtilis in Escherichia Coli

FU Yu-qin, LI Min, YUE Xiao-jing, CUI Yin-qiu, ZHOU Hui

(Macromolecular Laboratory, College of Life Science, Jilin University, Changchun 130021, China)

Received:2005-08-26 Revised:1900-01-01 Online:2006-03-26 Published:2006-03-26
Contact: CUI Yin-qiu

摘要/Abstract

摘要： 扩增了枯草芽孢杆菌的ade基因，重组入载体pMal-c2x中，构建了麦芽糖结合蛋白（MBP）融合蛋白的表达体系. 通过IPTG诱导表达，用MBP亲和层析法，纯化该融合蛋白（104 700），并通过SDS-PAGE对表达及纯化结果进行检验，对其酶学性质进行了初步研究. 分析结果表明: 该融合酶蛋白具有显著的腺嘌呤脱氨酶活性，证明了ade基因是枯草芽孢杆菌中编码腺嘌呤脱氨酶的基因.

关键词: 腺嘌呤脱氨酶, MBP融合表达, 活性测定

Abstract: The ade gene, which was presumed to encode adenine deaminase in Bacillus subtilis, was cloned, then the gene was inserted into expression vector pMal-c2x and transformed into Escherichia coli TB1. The fusion expression of the ade gene product with MBP was induced by IPTG. After purification by MBP affinity chromatography, the fusion protein was detected through SDS-PAGE. The deaminase activity of the fusion protein and related enzyme properties also were analyzed. The results show that the fusion protein had significant deaminase activity and verify that the ade gene was the coding gene of adenine deaminase. The study is very important for us to investigate the metabolism of adenine in Bacillus subtilis.

Key words: adenine deaminase, MBP fusion expression, activity measurement

中图分类号:

Q783

付玉芹, 李敏, 岳晓婧, 崔银秋, 周慧. 枯草芽孢杆菌的ade基因在大肠杆菌中的MBP融合表达及活性鉴定[J]. J4, 2006, 44(02): 299-302.

FU Yu-qin, LI Min, YUE Xiao-jing, CUI Yin-qiu, ZHOU Hui. Fusion Expression and Characterization of the ade Gene from Bacillus Subtilis in Escherichia Coli[J]. J4, 2006, 44(02): 299-302.