title: Cloning and molecular characterisation of resuscitation promoting factor-like gene from Mycobacterium avium subspecies avium.
abstract: OBJECTIVE
  Resuscitation promoting factor (Rpf)-like gene of Mycobacterium avium subspecies paratuberculosis has been known to stimulate the growth of mycobacteria and enhances the recovery of replicating cells from non-replicating phases. The objective of the study was to produce recombinant rpf-like protein of M. avium subspecies avium protein for purification and physico-chemical characterisation.
  
  
  METHODS
  The identified rpf gene of M. avium subspecies avium was cloned, subcloned, sequenced and expressed in Escherichia coli expression system for the production of the recombinant protein. The expressed recombinant Rpf protein was confirmed by Western blot and the extract was purified to yield a pure recombinant protein.
  
  
  RESULTS
  An rpf-like gene of 675 bp size in the M. avium subspecies avium was identified. This gene was expressed and the recombinant Rpf weighed 65 kDa as confirmed by Western blot. The M. avium recombinant Rpf protein was extracted under denatured conditions and purified yielding a recombinant protein with >90% purity.
  
  
  CONCLUSIONS
  Identification, cloning, sequencing and expression of a rpf-like gene from M. avium suggest that RpfA is present in this species also, which might be involved in reactivation phenomenon in this high-risk pathogen.
authors.forename: R
authors.initials: R
authors.lastname: Kavitha
authors.affiliation: Bacteriology and Mycology Division, Mycobacteria Laboratory, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, india.
authors.forename: R
authors.initials: R
authors.lastname: Verma
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D015153:Blotting, Western; D003001:Cloning, Molecular; D016207:Cytokines / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D008970:Molecular Weight; D009162:Mycobacterium avium / Q000235:genetics*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D016207:Cytokines; D011994:Recombinant Proteins; C113704:resuscitation-promoting factor, bacteria