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在大肠杆菌中生产重组丝氨酸蛋白酶抑制剂

Production of recombinant serpins in Escherichia coli.

作者信息

Pearce Mary C, Cabrita Lisa D

机构信息

Department of Biochemistry, Monash University, Clayton, Australia.

出版信息

Methods Enzymol. 2011;501:13-28. doi: 10.1016/B978-0-12-385950-1.00002-X.

Abstract

Serpins represent a diverse family of proteins that are found in a wide range of organisms and cellular locations. In order to study them, most need to be produced recombinantly, as isolation from their source is not always possible. Due to their relatively uncomplicated structure (single domain, few posttranslational modifications), the serpins are usually amenable to expression in Escherichia coli, which offers a fast and cost-effective solution for the generation of large amounts of protein. This chapter outlines the general procedures used in the expression and subsequent purification of serpins in E. coli, with a particular focus on the methods used for antitrypsin, the archetypal member of the family.

摘要

丝氨酸蛋白酶抑制剂是一类多样的蛋白质家族,存在于广泛的生物体和细胞位置中。为了研究它们,大多数需要通过重组方式生产,因为从其来源进行分离并不总是可行的。由于其相对简单的结构(单结构域,很少有翻译后修饰),丝氨酸蛋白酶抑制剂通常适合在大肠杆菌中表达,这为大量蛋白质的产生提供了一种快速且经济高效的解决方案。本章概述了在大肠杆菌中表达并随后纯化丝氨酸蛋白酶抑制剂所使用的一般程序,特别关注用于该家族原型成员抗胰蛋白酶的方法。

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