Tanimoto K, Tomita H, Ike Y
Department of Microbiology, Gunma University School of Medicine, Japan.
Plasmid. 1996 Jul;36(1):55-61. doi: 10.1006/plas.1996.0032.
The Enterococcus faecalis conjugative plasmid pPD1 (59 kb) produces an aggregation substance in response to pheromone. The traA gene (962 bp) is a member of a gene cluster involved in the regulation of the pPD1 pheromone response. A chimeric E. faecalis suicide plasmid containing a 762-bp DNA fragment from the middle portion of the traA gene was constructed. The pPD1 traA was disrupted by integration of this chimeric plasmid via homologous recombination. The E. faecalis strain containing pPD1 with the disrupted traA exhibited dry colony morphology, constitutive clumping, and an ability to transfer at high frequencies in a short (10-min) mating period, indicating that the traA product of pPD1 is a negative regulator for the pheromone response.
粪肠球菌接合质粒pPD1(59 kb)在信息素的作用下产生聚集物质。traA基因(962 bp)是参与pPD1信息素应答调控的基因簇成员。构建了一个嵌合的粪肠球菌自杀质粒,其包含来自traA基因中部的762 bp DNA片段。通过同源重组,该嵌合质粒的整合破坏了pPD1的traA。含有被破坏traA的pPD1的粪肠球菌菌株表现出干燥菌落形态、组成型聚集,并且能够在短(10分钟)交配期内高频转移,这表明pPD1的traA产物是信息素应答的负调节因子。