Factors required in vitro for excision of the Bacteroides conjugative transposon, CTnDOT

Plasmid
Y SutantoA A Salyers

Abstract

Four genes have been found to be essential for excision of the Bacteroides conjugative transposon CTnDOT in vivo: intDOT, orf2c, orf2d, and exc. The intDOT gene encodes an integrase that is essential for integration and excision. The function of the other genes is still uncertain. Previously, we developed an in vitro system for the integration reaction. We have now developed an in vitro system for excision. In this system, the left and right junctions of CTnDOT, attL, and attR, are provided on separate plasmids. The excision reaction produced a cointegrate which contained the attDOT (the joined ends of CTnDOT) and attB (the chromosomal target site). Cointegrate formation was observed after electroporation of Escherichia coli with the assay mixture and was also detected directly in the assay mixture by Southern hybridization. The highest reaction frequencies (10(-3)) were obtained with a mixture that contained purified IntDOT and a cell extract from Bacteroides thetaiotaomicron 4001, which contained the excision region of CTnDOT carried on a plasmid. An unexpected finding was that the addition of purified Exc, which is essential for excision in vivo, was not required for excision in vitro, nor did it increase the frequency of co...Continue Reading

References

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Citations

Apr 17, 2012·Journal of Bacteriology·Carolyn M Keeton, Jeffrey F Gardner
Jul 3, 2007·Journal of Bacteriology·Asa Flanigan, Jeffrey F Gardner
Nov 17, 2009·Journal of Bacteriology·Seyeun KimJeffrey F Gardner
Feb 6, 2007·Journal of Bacteriology·Jeanne M DichiaraJeffrey F Gardner
Aug 4, 2005·Journal of Bacteriology·Kyung MoonAbigail A Salyers
Apr 5, 2005·Journal of Bacteriology·M BacicC J Smith
Jun 8, 2006·Annual Review of Biochemistry·Nigel D F GrindleyPhoebe A Rice
May 13, 2006·Molecular Microbiology·Karolina MalanowskaJeffrey F Gardner
Apr 23, 2013·Plasmid·Carolyn M KeetonJeffrey F Gardner
Mar 1, 2017·Journal of Bacteriology·Kenneth RingwaldJeffrey Gardner
May 11, 2007·FEMS Microbiology Letters·Rustam I Aminov, Roderick I Mackie
Apr 4, 2017·FEMS Microbiology Reviews·François DelavatJan Roelof van der Meer
Jun 25, 2015·Microbiology Spectrum·Margaret M Wood, Jeffrey F Gardner

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