枯草芽孢杆菌HPB12-L24核糖体蛋白的过量生产、纯化及特性分析

Overproduction, purification and characterization of the HPB12-L24 ribosomal protein of Bacillus subtilis.

作者信息

Zouine M, Beloin C, Deneubourg A M, Hirschbein L, Le Hegarat F

机构信息

Institut de Génétique et Microbiologie, Université Paris-Sud, Orsay, France.

出版信息

FEMS Microbiol Lett. 1996 Nov 15;145(1):41-8. doi: 10.1111/j.1574-6968.1996.tb08554.x.

DOI:10.1111/j.1574-6968.1996.tb08554.x

PMID:8931325

Abstract

HPB12-L24 was previously described as a bifunctional histone-like and ribosomal protein in Bacillus subtilis. In order to confirm the identity of HPB12 and L24, and to study the properties of this protein, the rplX gene of B. subtilis encoding L24 has been overexpressed in Escherichia coli by an efficient protein overproduction system. A simple and rapid purification scheme using ammonium sulfate precipitation and cation-exchange chromatography is presented. 10 mg of pure L24 per g of Escherichia coli cells were obtained. The purified recombinant protein L24 is heat-stable, acid-soluble and binds preferentially supercoiled DNA like protein HPB12. These results confirm the identity of HPB12 and L24. Overexpression of rplX led to gross alterations of cell morphology and to an abnormal shape of nucleoids.

摘要

HPB12-L24先前被描述为枯草芽孢杆菌中的一种双功能组蛋白样和核糖体蛋白。为了确认HPB12和L24的同一性，并研究该蛋白的特性，通过高效蛋白质过量表达系统在大肠杆菌中过表达了枯草芽孢杆菌编码L24的rplX基因。本文介绍了一种使用硫酸铵沉淀和阳离子交换色谱的简单快速纯化方案。每克大肠杆菌细胞可获得10毫克纯L24。纯化的重组蛋白L24具有热稳定性、酸溶性，并且像蛋白HPB12一样优先结合超螺旋DNA。这些结果证实了HPB12和L24的同一性。rplX的过表达导致细胞形态的显著改变和类核形状异常。