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液化沙雷氏菌作为一种新型宿主,可利用大肠杆菌的N-乙酰神经氨酸裂解酶基因进行过量生产和纯化。

Serratia liquefaciens as a new host superior for overproduction and purification using the N-acetylneuraminate lyase gene of Escherichia coli.

作者信息

Yamamoto K, Kawakami B, Kawamura Y, Kawai K

机构信息

Tsuruga Institute of Biotechnology, Toyobo Co., Ltd., Fukui, Japan.

出版信息

Anal Biochem. 1997 Mar 15;246(2):171-5. doi: 10.1006/abio.1997.2001.

Abstract

Serratia liquefaciens was screened as a host strain for effective gene expression and easy purification of the target protein. A model gene, N-acetylneuraminate lyase gene (nanA), fused with the promoter region of Escherichia coli lac operon successfully overproduced the protein independently from the inducer. Since S. liquefaciens grew at lower temperature than E. coli and its proteins were more heat sensitive than those of E. coli, simple incubation at 60 degrees C could inactivate most enzymes but the nanA protein. Subsequent column works for purification, then, became simple and rapid.

摘要

粘质沙雷氏菌被筛选作为宿主菌株,用于有效基因表达和目标蛋白的简易纯化。一个模型基因,即N - 乙酰神经氨酸裂解酶基因(nanA),与大肠杆菌乳糖操纵子的启动子区域融合,成功地独立于诱导剂过量表达了该蛋白。由于粘质沙雷氏菌在比大肠杆菌更低的温度下生长,且其蛋白质比大肠杆菌的蛋白质对热更敏感,因此在60摄氏度简单孵育就能使大多数酶失活,但nanA蛋白不受影响。随后的纯化柱操作就变得简单而快速。

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