[1]季勤,张云峰,罗玉明,等.高盐胁迫渗透对SBD2重组蛋白在大肠杆菌中可溶性表达的影响[J].南京师大学报(自然科学版),2010,33(03):70-75.
　Ji Qin,Zhang Yunfeng,Luo Yuming,et al.Effect of Hyperosmotic Stress on Expression of Soluble SBD2 Recombinant Protien in Escherichia coli[J].Journal of Nanjing Normal University(Natural Science Edition),2010,33(03):70-75.
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高盐胁迫渗透对SBD2重组蛋白在大肠杆菌中可溶性表达的影响()
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《南京师大学报（自然科学版）》[ISSN:1001-4616/CN:32-1239/N]

卷:
第33卷
期数:
2010年03期
页码:
70-75
栏目:
生命科学
出版日期:
2010-09-20

文章信息/Info

Title:
Effect of Hyperosmotic Stress on Expression of Soluble SBD2 Recombinant Protien in Escherichia coli
作者:
季勤1;  张云峰1;  罗玉明1;  徐春1;  黄翠香2
1. 淮阴师范学院生命科学学院, 江苏省环洪泽湖生态农业生物技术重点实验室, 江苏淮安223300 2. 南京师范大学生命科学学院, 生物多样性与生物技术江苏省重点实验室, 江苏南京210046
Author(s):
Ji Qin1; Zhang Yunfeng1; Luo Yuming1; Xu Chun1; Huang Cuixiang2
1.Jiangsu Key Laboratory for Eco-Agricultural and Biotechnology Around Hongze Lake,School of Life Sciences,Huaiyin Normal University,Huaian 223300,China 2. Jiangsu Key Laboratory for B iodivers ity and B iotechnology, Schoo l ofL ife Scien ce, Nan j ing N
关键词:
SBD重组蛋白;  相容性溶质;  高渗胁迫;  可溶性表达
Keywords:
SBD2 pro te in;  compatib le solute;  hyperosmo tic stress;  so luble expression
分类号:
Q786
摘要:
在基因工程的操作中,包涵体的形成是人们利用大肠杆菌表达外源蛋白质的一大难点.本研究将环状芽孢杆菌(Bacillus circulans)环状糊精糖基转移酶(CGTase)的淀粉粒结合域(SBD)基因编码序列的两个拷贝通过一连接肽连接(SBD2)后,克隆到大肠杆菌表达载体pTrcHis B上,得到的pTrcHis B/SBD2质粒转化大肠杆菌Top10,研究了不同培养基、不同诱导温度和时间对SBD2蛋白表达的影响.结果表明,利用相容性溶质山梨醇和甜菜碱等,在高盐胁迫下,实现了SBD2重组蛋白质在大肠杆菌中的
Abstract:
An artificial tandm repeat o f a starch-binding dom a in ( SBD2) was eng ineered by tw o copies o f the SBD derived from Bacillus circulans cyc lodex tr in g ly cosy ltransfe rase v ia a linke r peptide. The SBD2 gene was inserted into an E. coli expression v ector, pTrcH is B, to obtain the pTrcH is B /SBD2 vec to r. Recomb inantE. coli Top 10 ce lls, conta-i n ing the pTrcH is B /SBD2construc t, w ere g rown in LB w ith 50μ g /mL am picillin and LB w ith 50μg /mL amp icillin, 1 mo l/L sorb itol and 2.5 mm o l/L g lycy l beta ine, respective ly. Express ion o f SBD2 pro te in w as induced by add ing o f IPTG and ana lyzed by SDS-PAGE. The results show that la rge am ounts of so luble pro tein w ere obta ined by g row ing and induc ing the cells under osmo tic stress in the presence o f so rb ito l and g lycy l beta ine.

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备注/Memo

备注/Memo:
基金项目:国家自然科学基金(30771367);江苏省高校高新技术产业发展基金(JHB04-043);淮阴师范学院教授基金(06HSJS025);“青蓝工程” 通讯联系人: 季勤, 博士, 教授, 研究方向: 分子生物学. E-mail:jiqin@ hytc. edu. cn
更新日期/Last Update: 2013-04-08