铜绿假单胞菌鞭毛运动相关基因的研究

建立优化的转化条件,将M u转座复合物电转化到临床分离的一株铜绿假单胞菌(P seud om onasaerug inosa)PA 68中,最高转化效率达3.66×104CFU/μg DNA.通过表型筛选,得到三株鞭毛运动能力缺陷的突变子,Sourn thern杂交证实转座子为单点插入.经基因克隆、核苷酸测序研究,证明转座子分别插入到uvrD、phzF 1、zw f三个基因中,这是首次在国际上将M u转座重组技术应用到鞭毛运动相关基因的研究中.由于人工M u转座技术具有随机单点插入的优点,克服了传统转座子能在染色体上迁移的缺点,为进一步研究P.aerug inosa的鞭毛运动机理及致病性奠定基础.

The Study of Genes Involved in Swimming Motility of Pseudomonas aeruginosa

In this study,Mu transposition recombination technique was used to study a cluster of genes required for swimming motility mediated by flagellum of P.aeruginosa.Optimum conditions including growth state of the strain,electroshock voltage were studied for the electroporation of a clinical isolate strain PA68 with Mu transposonase complexes,and get highest transformation efficiency.Three swimming motility deficient mutants were isolated.Mini-Mu transposon inserted into genomic DNA in a single copy,which was confirmed by Southern blotting.Gene cloning and sequencing of the region flanking the insertion revealed that mini-Mu integrated into gene uvrD,zwf and phzF1 respectively. Mu transposon recombination technique is a new efficient method to study the function of bacterial genome.It's first time to be utilized to study the genes involved in swimming motility of P.aeruginosa.