title: Towards the high-throughput expression of metalloproteins from the Mycobacterium tuberculosis genome.
abstract: The provision of high-quality protein in adequate quantities is a prerequisite for any structural genomics programme. A number of proteins from the Mycobacterium tuberculosis genome have been expressed and the success at each stage of the process assessed. Major difficulties have been encountered in the purification and solubilization of many of these proteins, most likely as a result of mis-folding. Some improvements have been made to the protocol but the overall success rate is still limited; however, the use of a cell-free protein expression system will circumvent some of the difficulties encountered. Alternative purification systems are also required and the properties of a mutant blue copper protein are described, that may offer a combined purification and tagging system.
authors.affiliation: Cell Signalling Laboratory, De Montfort University, Leicester LE1 9BH, UK. jfhall@dmu.ac.uk
authors.forename: John F
authors.initials: JF
authors.lastname: Hall
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Ellis
authors.forename: Takanori
authors.initials: T
authors.lastname: Kigawa
authors.forename: Takashi
authors.initials: T
authors.lastname: Yabuki
authors.forename: Takayoshi
authors.initials: T
authors.lastname: Matsuda
authors.forename: Eiko
authors.initials: E
authors.lastname: Seki
authors.forename: S Samar
authors.initials: SS
authors.lastname: Hasnain
authors.forename: Shigeyuki
authors.initials: S
authors.lastname: Yokoyama
mesh_terms: D002474:Cell-Free System; D003001:Cloning, Molecular / Q000379:methods*; D016680:Genome, Bacterial; D008667:Metalloproteins / Q000737:chemistry* / Q000235:genetics*; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011487:Protein Conformation; D040901:Proteomics
chemical_list: D008667:Metalloproteins