Construction of Mutation Library in Burkholderia cepacia byInserting Tn5 Transposon and the Screening of Relevant Strain

Chinese Agricultural Science Bulletin ›› 2014, Vol. 30 ›› Issue (9) : 242-247. DOI: 10.11924/j.issn.1000-6850.2013-1710
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Construction of Mutation Library in Burkholderia cepacia byInserting Tn5 Transposon and the Screening of Relevant Strain

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The plasmid pRLl063a containing Tn5 transposon was transferred into Burkholderia cepacia strains T1828 and ZWL15 by conjunction, and associators were screened on restrictive plates with kanamycin and strepmycin. 300 mutants were obtained by Tn5 randomly inserted. PCR test were positive for six randomly selected mutants, and the mutants could not synthesize coronatine by HPLC detection. The results showed that, the genome for six randomly selected mutants had been inserted by Tn5 and lose the ability to synthesize coronatine. This study provided a basis for cloning and sequencing the transposon insertion sites’flanking sequence and the recovery test function.

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Burkholderia cepacia; coronatine; Tn5 transposon; mutation library

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Construction of Mutation Library in Burkholderia cepacia byInserting Tn5 Transposon and the Screening of Relevant Strain. Chinese Agricultural Science Bulletin. 2014, 30(9): 242-247 https://doi.org/10.11924/j.issn.1000-6850.2013-1710

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