Enhanced expression of recombinant beta toxin of Clostridium perfringens type B using a commercially available Escherichia coli strain.
Enhanced expression of recombinant beta toxin of Clostridium perfringens type B using a commercially available Escherichia coli strain.
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DOI：
10.4102/ojvr.v83i1.1136
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发表时间：
2016-06-30
期刊：
影响因子：
--
通讯作者：
Eimani BG
中科院分区：
文献类型：
--
作者：
Bakhshi F;Pilehchian Langroudi R;Eimani BG
产气荚膜梭菌β毒素仅由B型和C型产生，在许多人类和动物疾病中起重要作用，导致源自肠道的致命疾病。我们比较了C.大肠杆菌菌株RosettaTM（DE 3）和BL 21（DE 3）中的产气荚膜梭菌B型疫苗株重组β毒素基因。从pJETβ中提取β毒素基因，与pET 22 b（+）连接。pET 22 β转化E. coli BL 21（DE 3）和RosettaTM（DE 3）。重组蛋白经异丙基β-D-1-硫代半乳糖苷（IPTG）诱导后在RosettaTM（DE 3）中以可溶性形式表达，而在BL 21（DE 3）中未表达。通过在37 °C和在0.5mM、ImM、1.5mM IPTG的诱导下生长重组细胞来优化表达。使用十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）评价表达。通过Ni-NTA纯化重组蛋白，并使用western blot进行分析。我们的结论是E. coli菌株RosettaTM（DE 3）能提高C.产气荚膜杆菌重组β毒素。
Clostridium perfringens beta toxin is only produced by types B and C and plays an important role in many human and animal diseases, causing fatal conditions that originate in the intestines. We compared the expression of C. perfringens type B vaccine strain recombinant beta toxin gene in the Escherichia coli strains RosettaTM(DE3) and BL21(DE3). The beta toxin gene was extracted from pJETβ and ligated with pET22b(+). pET22β was transformed into E. coli strains BL21(DE3) and RosettaTM(DE3). Recombinant protein was expressed as a soluble protein after isopropyl β-D-1-thiogalactopyranoside (IPTG) induction in strain RosettaTM(DE3) but not in BL21(DE3). Expression was optimised by growing recombinant cells at 37 °C and at an induction of 0.5 mM, 1 mM, 1.5 mM IPTG. Expression was evaluated using sodium dodecyl sulfate Polyacrylamide gel electrophoresis (SDS-PAGE). The recombinant protein was purified via Ni-NTA and was analysed using western blot. We concluded that E. coli strain RosettaTM(DE3) can enhance the expression of C. perfringens recombinant beta toxin.
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DOI：
10.1006/bbrc.1996.1422
发表时间：
1996-09-24
影响因子：
3.1
作者：
Goswami, PP;Rupa, P;Garg, LC
通讯作者：
Garg, LC
影响因子：
4.1
作者：
Nijland, Reindert;Lindner, Cordula;Kuipers, Oscar P.
通讯作者：
Kuipers, Oscar P.
影响因子：
3.6
作者：
CANARD, B;SAINTJOANIS, B;COLE, ST
通讯作者：
COLE, ST
影响因子：
2.1
作者：
STEINPORSDOTTIR, V;FRIDRIKSDOTTIR, V;ANDRESSON, OS
通讯作者：
ANDRESSON, OS
影响因子：
4.8
作者：
Nagahama, M;Hayashi, S;Sakurai, J
通讯作者：
Sakurai, J