1887

Abstract

The mechanisms that promote and regulate transcription in mycoplasmas are poorly understood. Here, a promoter-probe vector based on the pMTn minitransposon and containing a promoterless reporter gene was constructed to analyse transcription . Recovered transposon insertions were in monocopy, with 16 % expressing enough -galactosidase (-Gal) to yield colonies exhibiting a detectable blue colour. A sample of 52 blue colonies was propagated and selected for further analyses. The -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 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 .

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2007-08-01
2019-10-23
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Transposon insertions found inside coding regions [ PDF] (12 kb) Transposon insertion sites found in the genome. The red downward-facing triangles in the upper line indicate transposon insertion sites in the plus strand; the blue upward-facing triangles indicate transposon insertion sites insertion in the minus strand. The transcription terminator sequences were determined by the TransTerm program (Ermolaeva , 2000) and are marked by a T facing up (terminator sequence in the plus strand) or a T facing down (terminator sequence in the minus strand). A "T" symbol in red indicates the terminator sequence determined experimentally by Benders (2005). MgPa islands I to VIII contain sequences derived from mg191 and mg192 ORFs. The MgPa191 island exhibits only a short stretch of a sequence related to the mg191 ORF. [ PDF] (2245 kb) BLAST alignment of sequences in MgPa island VI surrounding the transcriptional start point and mg192 gene in the MgPa operon. The sequences in the alignment are from the minus strand of the genome and are numbered starting at base 314,465 (MgPa island VI) or 226,395 (mg192) of the complete genome. The heterogeneous transcriptional start point is boxed in green and the -10 region of a putative promoter, in yellow. [ PDF] (572 kb) Transcriptional analysis of the conserved gene cluster in and . , 4542-4551. Prediction of transcription terminators in bacterial genomes. , 27-33.

PDF

Transposon insertions found inside coding regions [ PDF] (12 kb) Transposon insertion sites found in the genome. The red downward-facing triangles in the upper line indicate transposon insertion sites in the plus strand; the blue upward-facing triangles indicate transposon insertion sites insertion in the minus strand. The transcription terminator sequences were determined by the TransTerm program (Ermolaeva , 2000) and are marked by a T facing up (terminator sequence in the plus strand) or a T facing down (terminator sequence in the minus strand). A "T" symbol in red indicates the terminator sequence determined experimentally by Benders (2005). MgPa islands I to VIII contain sequences derived from mg191 and mg192 ORFs. The MgPa191 island exhibits only a short stretch of a sequence related to the mg191 ORF. [ PDF] (2245 kb) BLAST alignment of sequences in MgPa island VI surrounding the transcriptional start point and mg192 gene in the MgPa operon. The sequences in the alignment are from the minus strand of the genome and are numbered starting at base 314,465 (MgPa island VI) or 226,395 (mg192) of the complete genome. The heterogeneous transcriptional start point is boxed in green and the -10 region of a putative promoter, in yellow. [ PDF] (572 kb) Transcriptional analysis of the conserved gene cluster in and . , 4542-4551. Prediction of transcription terminators in bacterial genomes. , 27-33.

PDF

Transposon insertions found inside coding regions [ PDF] (12 kb) Transposon insertion sites found in the genome. The red downward-facing triangles in the upper line indicate transposon insertion sites in the plus strand; the blue upward-facing triangles indicate transposon insertion sites insertion in the minus strand. The transcription terminator sequences were determined by the TransTerm program (Ermolaeva , 2000) and are marked by a T facing up (terminator sequence in the plus strand) or a T facing down (terminator sequence in the minus strand). A "T" symbol in red indicates the terminator sequence determined experimentally by Benders (2005). MgPa islands I to VIII contain sequences derived from mg191 and mg192 ORFs. The MgPa191 island exhibits only a short stretch of a sequence related to the mg191 ORF. [ PDF] (2245 kb) BLAST alignment of sequences in MgPa island VI surrounding the transcriptional start point and mg192 gene in the MgPa operon. The sequences in the alignment are from the minus strand of the genome and are numbered starting at base 314,465 (MgPa island VI) or 226,395 (mg192) of the complete genome. The heterogeneous transcriptional start point is boxed in green and the -10 region of a putative promoter, in yellow. [ PDF] (572 kb) Transcriptional analysis of the conserved gene cluster in and . , 4542-4551. Prediction of transcription terminators in bacterial genomes. , 27-33.

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