转座子标签法突变呋喃丹降解菌CFDS-1  

Screening of Mutants of Carbofuran Degrading Bacterium CFDS-1 by Transposon Tagging

作　　者：徐剑宏[1] 洪青[1] 汪婷[1] 张小华[1] 李顺鹏[1] 

机构地区：[1]南京农业大学农业部农业环境微生物工程重点开放实验室,南京210095

出　　处：《微生物学通报》2005年第2期34-38,共5页Microbiology China

基　　金：国家自然科学基金(No.30400013);国家高技术研究发展计划项目(No.2003AA241150);江苏省科技攻关项目(No.BE2002345;BE2003343)

摘　　要：通过接合使供体大肠杆菌DH5α中的质粒pSC123上的转座子插入到受体菌CFDS1基因组DNA中,以引起该菌株的基因插入突变。利用转座子上的卡那霉素抗性基因和呋喃丹降解过程中红色物质的产生与否初步筛选出6株突变株,分别命名为CFDSM1～CFDSM6。紫外扫描和气谱检测结果进一步证明这些突变子确实失去了对呋喃丹的降解能力。根据转座子的序列设计引物,以6株突变株的基因组DNA为模板进行PCR扩增,并对PCR产物进行限制性酶切分析,结果表明这些突变子中呋喃丹降解基因的失活就是由于转座子的插入而导致的。In order to mutate a carbofuran degrading strain CFDS-1,transposon of pSC123 was introduced into the genomic DNA of strain CFDS-1 by conjugation with E.coli DH5α (pSC123) as donor and strain CFDS-1 as recipient,6 mutants which lost carbofuran degrading ability were obtained with a kanamycin resistant gene in the middle of transposon and the disappearance of a red compound during the degrading of carbofuran as preliminary selecting marks,they were designated as CFDS-M1～CFDS-M6. UV scanning and GS assay results also proved their mutations. PCR was carried out respectively with primers designed according to the sequence of transposon and genomic DNA of 6 mutants as templates,restriction analysis of PCR products showed that the mutation of carbofuran degrading genes of these mutants was caused by transposon insertion.

关 键 词：呋喃丹 转座子标签法 降解 突变 

分 类 号：Q933[生物学—微生物学]