Improved Antibiotic Resistance Gene Cassette for Marker Exchange Mutagenesis in Ralstonia solanacearum and Burkholderia Species
2011
Um, H.Y., Dong-A University, Busan, Republic of Korea | Chung, E.S., Dong-A University, Busan, Republic of Korea | Lee, J.H., Dong-A University, Busan, Republic of Korea | Lee, S.W., Dong-A University, Busan, Republic of Korea
Marker exchange mutagenesis is a fundamental approach to understanding gene function at a molecular level in bacteria. New plasmids carrying a kanamycin resistance gene or a trimethoprim resistance gene were constructed to provide antibiotic resistance cassettes for marker exchange mutagenesis in Ralstonia solanacearum and many antibiotic-resistant Burkholderia spp. Insertion sequences present in the flanking sequences of the antibiotic resistance cassette were removed to prevent aberrant gene replacement and polar mutation during mutagenesis in wild-type bacteria. Plasmids provided in this study would be convenient for use in gene cassettes for gene replacement in other Gram-negative bacteria.
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