Application of 2D-DIGE in Identification of Differentially Expressed Proteins of Mycobacterium Bovis BCG in Nonculturable and Rpf Mediated Resuscitation Phase

Authors:

Amita Yadav¹, Ravi Kr Gupta¹, A Srinivasan², Tej P Singh², Brahm S Srivastava¹ and Ranjana Srivastava^1*

DOI: 10.23880/oajprs-16000112

Volume 2, Issue 1

Abstract

Prolonged incubation of Mycobacterium tuberculosis and Mycobacterium bovis BCG culture in stationary phase induces a true dormancy generating nonculturable (NC) cells. Recovery of NC cells to replicating phase can be promoted by resuscitation promoting factor (Rpf) of Micrococcus luteus. There are five homologs of Rpf proteins in M. tuberculosis and M. bovis BCG and all are individually able to resuscitate the nonculturable cells of mycobacteria. In this study, we have exploited fluorescent 2D-DIGE technique to compare the protein expression profile in nonculturable and resuscitation phase cells of M. bovis BCG. The study led us to identify eight proteins that were abundantly expressed in resuscitation phase and eight proteins with pronounced expression in nonculturable state. The differential expression of corresponding genes in two phases of growth was also analyzed at mRNA level using real time-PCR analysis. The study suggests several differentially expressed proteins belonging to intermediary metabolism and respiration, lipid metabolism, information pathway, virulence detoxification and adaptation, cell wall and cell processes categories at protein as well as RNA level that intend their role in the resuscitation phase, which might be crucial in exit from dormancy.

Keywords:

2D-DIGE; Rpf; Real time-PCR; Differential proteome analysis; Dormancy

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