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转化生长因子α-铜绿假单胞菌外毒素融合蛋白TGFα-PE40高效表达质粒的构建与表达

Construction and expression of high efficiency expressing plasmid of transforming growth factor alpha-Pseudomonas aeruginosa exotoxin fusion protein TGF alpha-PE40.

作者信息

Xu Y, Shen M, Zhao Q, Zhang H, Xu Y

机构信息

Laboratory of Cellular and Molecular Oncology, Shanghai Institute of Cell Biology, Chinese Academy of Sciences, China.

出版信息

Chin J Biotechnol. 1994;10(4):233-9.

PMID:7780019
Abstract

The XbaI/EcoRI cleaved TGF alpha-PE40 gene from plasmid pXY382 was inserted into the same cloning site of the expression vector pCB604 resulting in the plasmid p2X-TP1. E. coli BL21 (lambda DE3) cells were transformed with the p2X-TP1 and then induced with IPTG. The product expressed was accumulated mainly in the form of inclusion bodies. The expression level was closely related to the cell density, induction temperature and medium but not to the inductor dosage and the induction period within certain range. The expressed amount of the fusion protein TGF alpha-PE40 was about 50 mg/L.

摘要

从质粒pXY382上切下的经XbaI/EcoRI酶切的TGFα-PE40基因被插入到表达载体pCB604的相同克隆位点,得到质粒p2X-TP1。用p2X-TP1转化大肠杆菌BL21(λDE3)细胞,然后用异丙基-β-D-硫代半乳糖苷(IPTG)诱导。表达产物主要以包涵体形式积累。表达水平与细胞密度、诱导温度和培养基密切相关,但在一定范围内与诱导剂用量和诱导时间无关。融合蛋白TGFα-PE40的表达量约为50mg/L。

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