A new promoterless reporter vector reveals antisense transcription in Mycoplasma genitalium

Maria Lluch-Senar, Miquel Vallmitjana, Enrique Querol, Jaume Piñol

Research output: Contribution to journalArticleResearchpeer-review

19 Citations (Scopus)

Abstract

The mechanisms that promote and regulate transcription in mycoplasmas are poorly understood. Here, a promoter-probe vector based on the pMTnTetM438 minitransposon and containing a promoterless lacZ reporter gene was constructed to analyse Mycoplasma genitalium transcription in vivo. Recovered transposon insertions were in monocopy, with 16% expressing enough β-galactosidase (β-Gal) to yield colonies exhibiting a detectable blue colour. A sample of 52 blue colonies was propagated and selected for further analyses. The β-Gal activity of the corresponding cultures was measured to quantify, in a reproducible way, the transcription levels of the interrupted ORFs. Several insertions were found in sense with the interrupted ORF, but surprisingly there was also a number of insertions in non-coding regions, many of them in repetitive DNA regions known as MgPa islands. Moreover, 30% of the analysed transposon insertions had the lacZ gene in the opposite orientation to the coding frame, suggesting the existence of antisense transcripts that may be involved in the control of gene expression in M. genitalium. © 2007 SGM.
Original languageEnglish
Pages (from-to)2743-2752
JournalMicrobiology
Volume153
Issue number8
DOIs
Publication statusPublished - 1 Aug 2007

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