Construction of a large-scale Burkholderia cenocepacia J2315 transposon mutant library
Type
Conference PaperKAUST Department
Biological and Environmental Sciences and Engineering (BESE) DivisionBioscience Program
Date
2014-09-04Online Publication Date
2014-09-04Print Publication Date
2014Permanent link to this record
http://hdl.handle.net/10754/652853
Metadata
Show full item recordAbstract
Burkholderia cenocepacia, a pathogenic member of the Burkholderia cepacia complex (Bcc), has emerged as a significant threat towards cystic fibrosis patients, where infection often leads to the fatal clinical manifestation known as cepacia syndrome. Many studies have investigated the pathogenicity of B. cenocepacia as well as its ability to become highly resistant towards many of the antibiotics currently in use. In addition, studies have also been undertaken to understand the pathogen's capacity to adapt and survive in a broad range of environments. Transposon based mutagenesis has been widely used in creating insertional knock-out mutants and coupled with recent advances in sequencing technology, robust tools to study gene function in a genome-wide manner have been developed based on the assembly of saturated transposon mutant libraries. In this study, we describe the construction of a large-scale library of B. cenocepacia transposon mutants. To create transposon mutants of B. cenocepacia strain J2315, electrocompetent bacteria were electrotransformed with the EZ-Tn5 <TET-1> transposome. Tetracyline resistant colonies were harvested off selective agar and pooled. Mutants were generated in multiple batches with each batch consisting of ~ 20,000 to 40,000 mutants. Transposon insertion was validated by PCR amplification of the transposon region. In conclusion, a saturated B. cenocepacia J2315 transposon mutant library with an estimated total number of 500,000 mutants was successfully constructed. This mutant library can now be further exploited as a genetic tool to assess the function of every gene in the genome, facilitating the discovery of genes important for bacterial survival and adaptation, as well as virulence.Citation
Wong Y-C, Pain A, Nathan S (2014) Construction of a large-scale Burkholderia cenocepacia J2315 transposon mutant library. Available: http://dx.doi.org/10.1063/1.4895267.Sponsors
This project was funded by the Ministry of Science, Technology and Innovation, Malaysia (MOSTI). Yee-Chin Wong was supported by the MyBrain15 scholarship from the Ministry of Higher Education Malaysia.Publisher
AIP PublishingConference/Event name
2014 Postgraduate Colloquium of the Universiti Kebangsaan Malaysia, Faculty of Science and Technology, UKM FST 2014Additional Links
https://aip.scitation.org/doi/abs/10.1063/1.4895267ae974a485f413a2113503eed53cd6c53
10.1063/1.4895267