Open AccessTowards the high‐throughput expression of metalloproteins from the Mycobacterium tuberculosis genome
Author(s) -
Hall John F.,
Ellis Mark J.,
Kigawa Takanori,
Yabuki Takashi,
Matsuda Takayoshi,
Seki Eiko,
Hasnain S. Samar,
Yokoyama Shigeyuki
Publication year - 2005
Publication title -
journal of synchrotron radiation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.172
H-Index - 99
ISSN - 1600-5775
DOI - 10.1107/s0909049504027864
Subject(s) - mycobacterium tuberculosis , genome , structural genomics , computational biology , mutant , protein expression , tuberculosis , mycobacterium , chemistry , biology , microbiology and biotechnology , biochemistry , genetics , gene , bacteria , protein structure , medicine , pathology
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.