Abstract:
:The first step in the transfer of the Bacteroides conjugative transposon CTnDOT is excision of the integrated element from the chromosome to form a circular transfer intermediate. Excision occurs only after the bacteria are exposed to tetracycline. Previously, four excision genes were identified. One was the integrase gene intDOT, which appeared to be expressed constitutively. Three other genes essential for excision (orf2c, orf2d, and exc) were found located in a cluster 13 kbp downstream of intDOT. By using uidA fusions and real-time reverse transcriptase PCR, we demonstrate here that the excision genes orf2c, orf2d, and exc are part of an operon that also contains open reading frame orf3, previously shown not to be essential for excision. We also show that operon expression is regulated at the transcriptional level in response to tetracycline. The transcript start site for the operon has been localized. Three CTnDOT regulatory genes are thought to be involved in tetracycline regulation of excision, rteA, rteB, and rteC. By placing rteC under the control of a heterologous promoter, we found that RteC alone was sufficient for induction of the orf2c operon. If, however, the rteC gene was under the control of its own promoter, it was not able to induce orf2c operon expression unless rteA and rteB were present. Thus, RteA and RteB participate in excision by stimulating transcription of rteC. Using electrophoretic mobility shift analysis, we found that a purified His(6)-tagged form of RteC bound DNA upstream of the -33 region of the promoter. Changing the sequence in the region between bp -50 and -70 reduced the expression of the orf2c operon in vivo. Taken together, our results support the hypothesis that RteC acts as a DNA-binding protein that binds upstream of the orf2c promoter and is responsible for tetracycline-regulated transcriptional regulation of the orf2c operon.
journal_name
J Bacterioljournal_title
Journal of bacteriologyauthors
Moon K,Shoemaker NB,Gardner JF,Salyers AAdoi
10.1128/JB.187.16.5732-5741.2005keywords:
subject
Has Abstractpub_date
2005-08-01 00:00:00pages
5732-41issue
16eissn
0021-9193issn
1098-5530pii
187/16/5732journal_volume
187pub_type
杂志文章abstract::Pertussis toxin (PT), the major virulence factor of Bordetella pertussis, is composed of five different subunits whose genes are organized as an operon. We report the mapping of the promoter region of the PT operon and show that this promoter is only weakly active in Escherichia coli. Bordetella parapertussis and Bord...
journal_title:Journal of bacteriology
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journal_title:Journal of bacteriology
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doi:10.1128/JB.112.3.1306-1309.1972
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更新日期:1990-03-01 00:00:00
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pub_type: 杂志文章
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journal_title:Journal of bacteriology
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.114.1.267-272.1973
更新日期:1973-04-01 00:00:00
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journal_title:Journal of bacteriology
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更新日期:1968-01-01 00:00:00
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更新日期:1971-04-01 00:00:00
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journal_title:Journal of bacteriology
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journal_title:Journal of bacteriology
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更新日期:2019-11-20 00:00:00
abstract::Two methods of mutagenesis, chemical alkylation and insertion of the tetracycline resistance transposon Tn10, were used to generate mutants of Salmonella typhimurium which had reduced levels of endodeoxyribonucleolytic activity. The chemically induced mutations defined a locus, endA, which was cotransduced with serA a...
journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:1980-10-01 00:00:00
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journal_title:Journal of bacteriology
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更新日期:1994-06-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.174.21.6896-6903.1992
更新日期:1992-11-01 00:00:00