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一种与细菌在巨噬细胞中存活相关的分枝杆菌蛋白的克隆、表达、纯化及初步X射线衍射研究

Cloning, expression, purification and preliminary X-ray diffraction studies of a mycobacterial protein implicated in bacterial survival in macrophages.

作者信息

Shahine Adam E, Chan Phooi Y, Littler Dene, Vivian Julian, Brammananth Rajini, Crellin Paul K, Coppel Ross L, Rossjohn Jamie, Beddoe Travis

机构信息

Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.

出版信息

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2013 May 1;69(Pt 5):566-9. doi: 10.1107/S1744309113010427. Epub 2013 Apr 30.

Abstract

Mycobacterium species have developed numerous strategies to avoid the antimycobacterial actions of macrophages, enabling them to survive within the generally inhospitable environment of the cell. The recently identified MSMEG_5817 protein from M. smegmatis is highly conserved in Mycobacterium spp. and is required for bacterial survival in macrophages. Here, the cloning, expression, purification and crystallization of MSMEG_5817 is reported. Crystals of MSMEG_5817 were grown in 1.42 M Li2SO4, 0.1 M Tris-HCl pH 7.7, 0.1 M sodium citrate tribasic dihydrate. Native and multiple-wavelength anomalous dispersion (MAD) data sets have been collected and structure determination is in progress.

摘要

分枝杆菌属已发展出多种策略来规避巨噬细胞的抗分枝杆菌作用,使其能够在细胞内通常不适宜生存的环境中存活。最近从耻垢分枝杆菌中鉴定出的MSMEG_5817蛋白在分枝杆菌属中高度保守,是细菌在巨噬细胞中存活所必需的。本文报道了MSMEG_5817的克隆、表达、纯化及结晶。MSMEG_5817晶体在1.42 M硫酸锂、0.1 M Tris-HCl pH 7.7、0.1 M 柠檬酸三钠二水合物中生长。已收集了天然和多波长反常散射(MAD)数据集,结构测定正在进行中。

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