文献类型：期刊文献

英文题名：[Cloning and expression of the genes of glutathione synthetases].

作者：Shen, L X[1];Wei, D Z;Zhao, Z F;Zhang, S L;Wang, E L

机构：[1]State Key Laboratory of Bioreactor Engineering, Institute of Biochemistry, East China University of Science & Technology, Shanghai 200237, China.

年份：2001

卷号：17

期号：1

外文期刊名：Sheng wu gong cheng xue bao = Chinese journal of biotechnology

收录：MEDLINE(收录号：11330198);;PubMed

语种：英文

摘要：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.