Journal of Microbiology

Research Support, Non-U.S. Gov'ts

NOTE] Improved Antibiotic Resistance Gene Cassette for Marker Exchange Mutagenesis in Ralstonia solanacearum and Burkholderia Species: Hae Young Um , Eunsook Chung , Jai-Heon Lee , Seon-Woo Lee; J. Microbiol. 2011;49(2):305-308. Published online May 3, 2011; DOI: https://doi.org/10.1007/s12275-011-0439-0

Abstract: 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.

Construction of an Escherichia-Pseudomonas Shuttle Vector Containing an Aminoglycoside Phosphotransferase Gene and a lacZ'''' Gene for α-Complementation: Bheong-Uk Lee , Ja-Heon Hong , Hyung-Yeel Kahng , Kye-Heon Oh; J. Microbiol. 2006;44(6):671-673.; DOI: https://doi.org/2458 [pii]

Abstract: A new 4.87 kb Escherichia-Pseudomonas shuttle vector has been constructed by inserting a 1.27 kb DNA fragment with a replication origin of a Pseudomonas plasmid pRO1614 into the 3.6 kb E. coli plasmid pBGS18. This vector, designated pJH1, contains an aminoglycoside phosphotransferase gene (aph) from Tn903, a lacZ'''' gene for α-complementation and a versatile multiple cloning site possessing unique restriction sites for EcoRI, SacI, KpnI, SmaI, BamHI, XbaI, SalI, BspMI, PstI, SphI, and HindIII. When pJH1 was transformed into E. coli DH5α and into P. putida HK-6, it was episomally and stably maintained in both strains. In addition, the enhanced green fluorescent protein (EGFP) gene which was transcriptionally cloned into pJH1 rendered E. coli cells fluorescence when its transformants were illuminated at 488 nm.

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