title: Cloning, expression, purification and preliminary X-ray diffraction studies of a mycobacterial protein implicated in bacterial survival in macrophages.
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.
authors.affiliation: Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.
authors.forename: Adam E
authors.initials: AE
authors.lastname: Shahine
authors.forename: Phooi Y
authors.initials: PY
authors.lastname: Chan
authors.forename: Dene
authors.initials: D
authors.lastname: Littler
authors.forename: Julian
authors.initials: J
authors.lastname: Vivian
authors.forename: Rajini
authors.initials: R
authors.lastname: Brammananth
authors.forename: Paul K
authors.initials: PK
authors.lastname: Crellin
authors.forename: Ross L
authors.initials: RL
authors.lastname: Coppel
authors.forename: Jamie
authors.initials: J
authors.lastname: Rossjohn
authors.forename: Travis
authors.initials: T
authors.lastname: Beddoe
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D003001:Cloning, Molecular / Q000379:methods; D015964:Gene Expression Regulation, Bacterial; D008264:Macrophages / Q000382:microbiology*; D020102:Mycobacterium smegmatis; D014961:X-Ray Diffraction
chemical_list: D001426:Bacterial Proteins
keywords: MSMEG_5817; Mycobacterium smegmatis; macrophages; selenomethionine