A new promoterless reporter vector reveals antisense transcription in Mycoplasma genitalium.

摘要: The mechanisms that promote and regulate transcription in mycoplasmas are poorly understood. Here, a promoter-probe vector based on the pMTnTetM438 minitransposon containing promoterless lacZ reporter gene was constructed to analyse Mycoplasma genitalium vivo. Recovered transposon insertions were monocopy, with 16 % expressing enough β-galactosidase (β-Gal) yield colonies exhibiting detectable blue colour. A sample of 52 propagated selected for further analyses. β-Gal activity corresponding cultures measured quantify, reproducible way, levels interrupted ORFs. Several found sense ORF, but surprisingly there also number non-coding regions, many them repetitive DNA regions known as MgPa islands. Moreover, 30 analysed had opposite orientation coding frame, suggesting existence antisense transcripts may be involved control expression M. genitalium.