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Genomics |
Immunopathology Unit1 and Protein Science Unit2, Glaxo Wellcome Research and Development, Medicines Research Centre, Stevenage, Herts, SG1 2NY, UK
Author for correspondence: Joanna C. Betts. Tel: +44 1438 768138. Fax: +44 1438 764898. e-mail: jb75084{at}glaxowellcome.co.uk
The genome sequences of two virulent strains of Mycobacterium tuberculosis (H37Rv and CDC 1551) are now available. CDC 1551 is a recent clinical isolate and H37Rv is a commonly used lab strain which has been subject to in vitro passage. The two strains have been shown to display differing phenotypes both in vivo and in vitro. The proteome of the two strains grown in liquid culture were examined over time to determine whether there are any major differences between them at the protein level and the differences were compared to the genome data. Total cell lysates of the two strains were analysed by two-dimensional electrophoresis. Approximately 1750 protein spots were visualized by silver staining and the protein profiles of the two strains were found to be highly similar. Out of a total of 17 protein spot differences, seven were unique to CDC 1551 and three to H37Rv. Two further spots showed increased intensity in H37Rv, one spot showed differing vertical mobility between the strains and four showed differing spot intensities with time. Twelve of the spot differences were identified using mass spectrometry; however, no obvious association with phenotype could be deduced. When genome differences were analysed and related to the proteome differences, a mobility shift identified in the MoxR protein could be explained by a point mutation at the gene level. This proteome analysis reveals that, despite having been maintained under vastly different conditions, namely in vitro passage and in vivo transmission, these two strains have remained highly similar.
Keywords: mycobacteria, tuberculosis, proteome, two-dimensional gel electrophoresis, genome
Abbreviations: 2D, two-dimensional; BCG, bacille Calmette–Guérin; IPG, immobilized pH gradient; MALDI, matrix-assisted laser desorption-ionization; PGRS, polymorphic G+C rich sequence; PE, Pro-Glu; PPE, Pro-Pro-Glu; TOF, time-of-flight
a Present address: Molecular Biology, AstraZeneca R&D Charnwood, Bakewell Road, Loughborough LE11 5RH, UK.
b Present address: Department of Molecular Biology and Microbiology, Tufts University, Boston, MA 02111, USA.
c Present address: Astex Technology, 250 Cambridge Science Park, Milton Road, Cambridge CB4 0WE, UK
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