Expression of a fusion protein of human proinsulin with glutathione-S-transferase in Escherichia coli

doi:10.4028/www.scientific.net/AMR.998-999.248

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	Expression of a fusion protein of human proinsulin with glutathione-S-transferase in Escherichia coli
	Di, Zhi Xin; Ma, Jian Zhong; Wang, Yong Gang
	2014
会议日期	July 28, 2014 - July 29, 2014
会议地点	Qingdao, China
关键词	Insulin Mass spectrometry Peptides Stars Urea E. coli BL21(DE3) Fusion proteins Glutathione-S-transferase Human proinsulin Inclusion bodies Mass spectra Sequence encoding Ultra-sonication
卷号	998-999
DOI	10.4028/www.scientific.net/AMR.998-999.248
页码	248-251
英文摘要	A DNA sequence encoding for the human proinsulin was designed according to the codon bias of Escherichia coli and then chemically synthesized. The synthesized DNA fragment was subcloned into pGEX-3X for expression in E. coli BL21 (DE3) and E. coli BL21 Star (DE3), respectively. Conditions for the highest expression of the GST-proinsulin fusion proteins were optimized. These conditions are that cells of E. coli BL21 star (DE3) are incubated in 100mL of the LB medium with 2 mmol/L IPTG and 60μg/mL ampicillin at 26°Cfor 4h. After disrupted E. coli cells with ultrasonication, inclusion bodies were precipitated from cell lysis and washed. Fusion proteins from the inclusion bodies were redissolved in 8mmol/L of urea. After dialysed in purified water, fusion proteins were analysed by SDS-PAGE. The purity of the fusion protein is about 80.5% in total. The fusion protein from SDS-PAGE was further identified by mass/mass spectrum. GST in the dyad protein is confirmed by the 9 matched sequences. However, the left part is proved a polypeptide of which is completely different from the human proinsulin. © (2014) Trans Tech Publications, Switzerland.
会议录	Advanced Materials Research
会议录出版者	Trans Tech Publications Ltd
语种	英语
ISSN号	10226680
内容类型	会议论文
源URL	[http://ir.lut.edu.cn/handle/2XXMBERH/117353]
专题	兰州理工大学生命科学与工程学院
作者单位	School of Life Science and Engineering, Lanzhou Univeristy of Technology, China
推荐引用方式 GB/T 7714	Di, Zhi Xin,Ma, Jian Zhong,Wang, Yong Gang. Expression of a fusion protein of human proinsulin with glutathione-S-transferase in Escherichia coli[C]. 见:. Qingdao, China. July 28, 2014 - July 29, 2014.