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Purification of MBP Fused Mycobacterium Tuberculosis Proteins
Author(s) -
Crenshaw Ezekiel Matthew David
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.498.1
Subject(s) - maltose binding protein , affinity chromatography , mycobacterium tuberculosis , tandem affinity purification , fusion protein , protein purification , flag tag , biochemistry , chemistry , mycobacterium smegmatis , structural genomics , biology , microbiology and biotechnology , protein structure , recombinant dna , tuberculosis , enzyme , gene , medicine , pathology
Mycobacterium tuberculosis (Mtb) is a bacterium that causes tuberculosis (TB), a bacterial infection that mainly affects the respiratory system. The objective of the TB Structural Genomics Consortium is to solve the structure of Mtb proteins related to disease, with the aim of determining the function associated with those structures which could potentially lead to new treatments. In order to solve the structure of those proteins, highly purified proteins are needed which is achieved through protein purification. Protein purification is important to determine the characteristics of the function, structure, and interactions of the protein of interest. Our method of purifying proteins is with a Maltose Binding Protein (MBP) which increases expression level and the solubility of our protein of interest. The first step is to express the fusion protein with MBP which is accomplished through fermentation. The next step is to isolate the fusion protein through His‐tag affinity chromatography. We then cleave the fusion protein with TEV Protease. After the cleavage has taken place, we remove the MBP using MBP affinity chromatography and/or Gel Filtration Chromatography is needed. In this poster, we show the process of isolating two Mtb target proteins,Rv0472 and Rv3567, fused with MBP.