Open AccessCloning, expression, purification and preliminary X‐ray diffraction studies of a mycobacterial protein implicated in bacterial survival in macrophages
Author(s) -
Shahine Adam E.,
Chan Phooi Y.,
Littler Dene,
Vivian Julian,
Brammananth Rajini,
Crellin Paul K.,
Coppel Ross L.,
Rossjohn Jamie,
Beddoe Travis
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309113010427
Subject(s) - cloning (programming) , protein expression , microbiology and biotechnology , diffraction , biology , chemistry , materials science , biochemistry , gene , physics , optics , computer science , programming language
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 Li 2 SO 4 , 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.