Genetic Dissection of a Prevalent Plasmid-Encoded Conjugation System in Lactococcus lactis.

Frontiers in Microbiology
Guillermo Ortiz CharnecoDouwe van Sinderen

Abstract

Plasmid pNP40, which was first identified nearly 40 years ago in Lactococcus lactis subsp. lactis biovar diacetylactis DRC3, encodes functions such as heavy metal-, bacteriophage-, and nisin-resistance, as well as plasmid transfer ability by conjugation. Here, we report an optimized conjugation protocol for this plasmid, yielding a transfer frequency that is approximately 4,000-fold higher than those previously reported in literature, while we also observed high-frequency plasmid co-mobilization. Individual mutations in 18 genes that encompass the presumed conjugation cluster of pNP40 were generated using ssDNA recombineering to evaluate the role of each gene in the conjugation process. A possible transcriptional repressor of this conjugation cluster, the product of the traR gene, was identified in this manner. This mutational analysis, paired with bioinformatic predictions as based on sequence and structural similarities, allowed us to generate a preliminary model of the pNP40 conjugation machinery.

References

Aug 1, 1992·PCR Methods and Applications·R S ChaW G Thilly
Oct 5, 1990·Journal of Molecular Biology·S F AltschulD J Lipman
Jan 1, 1969·Annual Review of Microbiology·R Curtiss
Apr 28, 1997·Molecular & General Genetics : MGG·E CabezónF de la Cruz
Feb 9, 2002·Journal of Clinical Laboratory Analysis·P MarkoulatosM Moncany
Mar 28, 2002·Nucleic Acids Research·A J EnrightC A Ouzounis
May 27, 2003·Trends in Biochemical Sciences·Alex Bateman, Neil D Rawlings
Jun 10, 2003·Microbiology and Molecular Biology Reviews : MMBR·Elisabeth GrohmannManuel Espinosa
Dec 19, 2003·Nucleic Acids Research·Alex BatemanSean R Eddy
Feb 21, 2004·FEMS Microbiology Reviews·M Victoria FranciaFernando de la Cruz
May 20, 2004·Journal of Bacteriology·Matthew C A Smith, Christopher D Thomas
Sep 4, 2004·Applied and Environmental Microbiology·Jonathan O'DriscollDouwe Van Sinderen
Jun 28, 2005·Nucleic Acids Research·Johannes SödingAndrei N Lupas
Feb 14, 2006·FEMS Microbiology Reviews·Susan MillsR Paul Ross
Sep 6, 2006·Journal of Bacteriology·Jonathan O'DriscollDouwe van Sinderen
Oct 13, 2006·Proceedings of the National Academy of Sciences of the United States of America·K MakarovaD Mills
Feb 3, 2009·Molecular Microbiology·Lorraine A DraperR Paul Ross
Dec 1, 2009·Microbiology and Molecular Biology Reviews : MMBR·Cristina E Alvarez-Martinez, Peter J Christie
Mar 10, 2010·BMC Bioinformatics·Doug HyattLoren J Hauser
Jul 21, 2010·Cellular Microbiology·Karin WalldenGabriel Waksman
Feb 14, 2012·Nucleic Acids Research·Jan-Peter van Pijkeren, Robert A Britton
Jun 1, 2013·Plasmid·Casey W Hemmis, Joel F Schildbach
May 9, 2014·Frontiers in Cellular and Infection Microbiology·Günther Koraimann, Maria A Wagner
May 28, 2014·FEMS Microbiology Reviews·Stuart AinsworthDouwe van Sinderen
Aug 27, 2014·FEMS Microbiology Reviews·Elena CabezónIgnacio Arechaga
Jun 25, 2015·Microbiology Spectrum·Nikolaus Goessweiner-MohrElisabeth Grohmann
Aug 27, 2016·Frontiers in Molecular Biosciences·Elisabeth GrohmannSabine Brantl
Dec 13, 2016·PloS One·Mariya TarazanovaHerwig Bachmann
Apr 6, 2017·Microbial Cell Factories·Adelene Ai-Lian SongRaha Abdul Rahim
Apr 26, 2017·Plasmid·Luis Alfredo Bañuelos-VazquezSusana Brom
Jun 9, 2017·PLoS Computational Biology·Ryan R WickKathryn E Holt
Sep 20, 2017·The EMBO Journal·Adam RedzejGabriel Waksman

❮ Previous
Next ❯

Software Mentioned

BLASTP
BLASTX
MCL
Artemis
HHPred
Prodigal
Pfam
SigmaPlot
TMHMM
SPSS

Related Concepts

Related Feeds

Bacteriophage: Phage Therapy

Phage therapy uses bacterial viruses (bacteriophages) to treat bacterial infections and is widely being recognized as an alternative to antibiotics. Here is the latest research.