%0 Journal Article
%T Cloning and Expression of the Genes of Glutathione Synthetases<br>谷胱甘肽合成酶系的克隆、测序及表达
%A L X Shen
%A D Z Wei
%A Z F Zhao
%A S L Zhang
%A E L Wang
%A <br>沈立新
%A 魏东芝
%A 赵哲峰
%A 张嗣良
%A 王二力
%J 生物工程学报
%D 2001
%I 
%X The genes(gsh-I,gsh-II) for gamma-glutamyl-cysteine synthetase(GSH-I) and glutathione synthetase(GSH-II) from Escherichia coli B were amplified by PCR and then subcloned into plasmid pUC19 respectively. The DNA fragments harboring gshII and gsh I were inserted into plasmid pTrc99A one by one to get a hybrid plasmid pTrc-gsh. E. coli BL21 was transformed by pTrc-gsh for expression of the related enzymes. Analysis of SDS-PAGE showed that the expected products were expressed. E. coli BL21(pTrc-gsh) were incubated at 37 degrees C and pH 7.2 to OD550 = 0.5. The conditions were then switched to 34 degrees C and pH6.7 after the addition of 0.1 mmol/L IPTG. The expressed products were up to 25% of the total protein of the bacteria. Acetone-treated cells of the engineered strain could synthesize GSH efficiently.
%K γ-谷氨酰半胱氨酸合成酶，谷胱甘肽合成酶
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=C1A4F9DD30724450&yid=14E7EF987E4155E6&vid=BCA2697F357F2001&iid=CA4FD0336C81A37A&sid=10F298ED9F164662&eid=8C83C265AD318E34&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=3&reference_num=9