Transposon mutagenesis of Psychrobacter cryohalolentis PAMC 21807 by tri-parental conjugation

• Main Authors: Jeong, Hyun-Jeong, Lee, Hyoungseok, Hong, Soon Gyu, Cho, Jang-Cheon, Lee, Hong Kum, Lee, Yoo Kyung
• Format: Article in Journal/Newspaper
• Language: English
• Published: Polar Research Institute of China - PRIC 2013
• Subjects: Fauna; Advances in Polar Science; Antarc*; Antarctic; King George Island; Polar Science
• Online Access: http://library.arcticportal.org/2502/; http://library.arcticportal.org/2502/1/A20130405.pdf

Random mutagenesis is commonly used to study gene function. The screening of mutants exhibiting specific phenotypes assists in the identification of phenotype-related genes. In the current study, we isolated Antarctic bacteria, and developed a transposon Tn5 mutagenesis system. A total of 26 strains were isolated from seawater and freshwater near Antarctic King Sejong Research Station, King George Island. Six Psychrobacter strains were identified as psychrophilic, with optimal growth temperatures of 10℃ or 15℃ Psychrobacter cryohalolentis PAMC 21807 with a high growth rate at 4℃ was selected for transposon mutagenesis. Tri-parental conjugation with a plasmid containing Tn5 produced 13 putative recombinants containing the selectable marker. Genomic Southern hybridization confirmed Tn5 existed as episomes for seven recombinants, and for a single recombinant, Tn5 was integrated into the genome of Psychrobacter cryohalolentis PAMC 21807. The result indicates that the mutagenesis method, although successful, has a relatively low rate. The psychrophilic bacteria isolated in this study may be a useful resource for studying cold adaptation mechanisms, and the mutagenesis method can be applied to genetic analysis.