R. Srivastava et al., Green fluorescent protein as a reporter in rapid screening of antituberculosis compounds in vitro and in macrophages, BIOC BIOP R, 253(2), 1998, pp. 431-436
Citations number
12
Categorie Soggetti
Biochemistry & Biophysics
Journal title
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
The development of new drugs against Mycobacterium tuberculosis is impeded
by slow growth and highly infectious nature of the organism that warrants t
he need to work under highly stringent biosafety conditions. These problems
can be overcome by use of reporter genes and surrogate strains. A strain o
f rapidly growing IM. aurum has been recommended as test organism to screen
inhibitors of mycobacteria to preselect compounds for progression into tes
ting against M. tuberculosis. We have investigated the application of recom
binant M. aurum expressing green fluorescent protein in rapid screening of
antituberculosis compounds in vitro and in infected macrophages. Recombinan
t M: aurum[pGFM-11] expressing green fluorescent protein was constructed. T
he assay is based on measurement of fluorescent intensity at 509 nm. A good
correlation was found between fluorescence and growth. Fluorescence of rec
ombinant IM. aurum was inhibited in vitro within 8 to 24 h by frontline ant
imycobacterial drugs at their reported MICs whereas inhibition in infected
macrophages was observed in 72 h. Therefore green fluorescent reporter syst
em provides a convenient screen to test antimycobacterial compounds that ar
e active in vitro and within infected macrophages. (C) 1998 Academic Press.