Towards the high-throughput expression of metalloproteins from the Mycobacterium tuberculosis genome

J Synchrotron Radiat. 2005 Jan;12(Pt 1):4-7. doi: 10.1107/S0909049504027864. Epub 2004 Dec 23.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell-Free System
  • Cloning, Molecular / methods*
  • Genome, Bacterial
  • Metalloproteins / chemistry*
  • Metalloproteins / genetics*
  • Mycobacterium tuberculosis / genetics
  • Mycobacterium tuberculosis / metabolism*
  • Polymerase Chain Reaction
  • Protein Conformation
  • Proteomics*

Substances

  • Metalloproteins