title: Cloning and expression of multiple integral membrane proteins from Mycobacterium tuberculosis in Escherichia coli.
abstract: Seventy integral membrane proteins from the Mycobacterium tuberculosis genome have been cloned and expressed in Escherichia coli. A combination of T7 promoter-based vectors with hexa-His affinity tags and BL21 E. coli strains with additional tRNA genes to supplement sparsely used E. coli codons have been most successful. The expressed proteins have a wide range of molecular weights and number of transmembrane helices. Expression of these proteins has been observed in the membrane and insoluble fraction of E. coli cell lysates and, in some cases, in the soluble fraction. The highest expression levels in the membrane fraction were restricted to a narrow range of molecular weights and relatively few transmembrane helices. In contrast, overexpression in insoluble aggregates was distributed over a broad range of molecular weights and number of transmembrane helices.
authors.affiliation: Department of Chemistry and Biochemistry, National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida 32306, USA.
authors.forename: Alla
authors.initials: A
authors.lastname: Korepanova
authors.forename: Fei P
authors.initials: FP
authors.lastname: Gao
authors.forename: Yuanzi
authors.initials: Y
authors.lastname: Hua
authors.forename: Huajun
authors.initials: H
authors.lastname: Qin
authors.forename: Robert K
authors.initials: RK
authors.lastname: Nakamoto
authors.forename: Timothy A
authors.initials: TA
authors.lastname: Cross
mesh_terms: D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D007425:Intracellular Membranes / Q000378:metabolism; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics*; D009169:Mycobacterium tuberculosis / Q000235:genetics*
chemical_list: D008565:Membrane Proteins