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

作　　者：Hyun-Jeong Jeong Hyoungseok Lee Soon Gyu Hong Jang-Cheon Cho Hong Kum Lee Yoo Kyung Lee 

机构地区：[1]Korea Polar Research Institute,KIOST,26 Songdomirae-ro,Yeonsu-gu,Incheon 406-840,Korea [2]Division of Biology and Ocean Sciences,Inha University,Incheon 402-751,Korea [3]Dongseo University & Techniche Universitt Berlin Joint Algae Laboratory (DUTUJAL),Division of Energy and Bioen-gineering,San 69-1 Churye-2-Dong,Sasang-Gu,Busan 617-716,Korea

出　　处：《Advances in Polar Science》2013年第4期223-230,共8页极地科学进展（英文版）

基　　金：supported by the Korea Polar Research Institute(Grant nos.PE08050 and PE13240)

摘　　要：Random mutagenesis is commonly used to study gene function. The screening of mutants exhibiting specific pheno- types 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 tempera- tures of 10~C or 15~C Psychrobacter cryohalolentis PAMC 21807 with a high growth rate at 4~C 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.Random mutagenesis is commonly used to study gene function. The screening of mutants exhibiting specific pheno- types 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 tempera- tures of 10~C or 15~C Psychrobacter cryohalolentis PAMC 21807 with a high growth rate at 4~C 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.

关 键 词：cold adaptation PSYCHROBACTER psychrophilic bacteria tri-parental conjugation transposon mutagenesis 

分 类 号：Q933[生物学—微生物学]