摘要
利用Tn5转座子插入突变技术构建沙雷氏Z4菌株突变体库,研究不同因素对转座突变的影响,在此基础上初步探究了突变株的群体淬灭能力.结果表明,采用双亲本接合法,选择对数生长期的沙雷氏Z4菌株与大肠杆菌S17-1(pRL1063a)菌液1∶1混合,30℃培养3 h后,涂布于含50μg/mL卡那霉素和25μg/mL四环素的LB固体培养基上,长出的突变株数量最多;随机选择突变株进行群体淬灭能力测试,结果筛选到了3株群体淬灭能力明显与沙雷氏Z4野生型有差异的突变株.本研究采用的Tn5转座突变技术为研究群体淬灭菌的基因功能以及生理代谢提供了有效的分子遗传工具,也为研究群体淬灭提供了新的途径.
In this study, the Tn5 transposon insertion mutation technique was used to construct the mutant library of the Sarratia Z4 strain, and the effects of different factors on transposable mutation were discussed.Based on this, the quorum quenching ability of the mutant was preliminarily investigated. The results showed that the number of mutants obtained by using the parent grafting method was the largest when the bacteria solution of E. coli S17-1(pRL1063a) and Sarratia sp. Z4 at logarithmic growth stage mixed 1∶1 was cultured at 30 ℃ for 3 h, and then coated on LB solid medium containing 50 μg/mL kanamycin and 25 μg/mL tetracycline. Mutant strains were selected randomly for quorum quenching ability test, and the quorum quenching ability of three mutant strains was significantly different from that of the wild type Sarratia sp. Z4.The Tn5 transposable mutation technique used in this study provided an effective molecular genetic tool for studying the gene function and physiological metabolism of quorum quenching sterilization, and also offered a new way for studying quorum quenching.
作者
杨惠婷
刘栋
赵一竹
杨瑞雪
葛世玫
YANG Huiting;LIU Dong;ZHAO Yizhu;YANG Ruixue;GE Shimei(College of Life and Environmental Science,Wenzhou University,Wenzhou,China 325035)
出处
《温州大学学报（自然科学版）》
2022年第2期44-53,共10页
Journal of Wenzhou University(Natural Science Edition)
基金
浙江省公益项目(LGF20E080022)
温州市优秀重点实验室基础性科研项目(H20210003)。
