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

与巨噬细胞中细菌存活有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

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

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.

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