鸭疫里默氏杆菌50S L27转座子插入突变株的生物学特性分析  

Partial Biological Characteristics of Transposon Insertion Mutant of 50S Ribosomal Protein L27 in Riemerelia Anatipestifer

作　　者：陈妍 丁云竹 杨永胜 徐欣欣 王西 宫艳杉 杜晓莉 常维山[3] 成大荣[1] 徐建生[1] 胡青海[2] CHEN Yan;DING Yunzhu;YANG Yongsheng;XU Xinxin;WANG Xi;GONG Yanshan;DU Xiaoli;CHANG Weishan;CHENG Darong;XU Jiansheng;HU Qinghai(College of Veterinary Medicine,Hangzhou University,Hangzhou 225009,China;Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China;College of Animal Science and Technology,Shandong Agricultural University,Taiwan 271018,China)

机构地区：[1]扬州大学兽医学院,扬州225009 [2]中国农业科学院上海兽医研究所,上海200241 [3]山东农业大学动物科技学院,泰安271018

出　　处：《中国动物传染病学报》2021年第1期21-25,共5页Chinese Journal of Animal Infectious Diseases

基　　金：国家自然科学基金(31772770,31472224)。

摘　　要：为了鉴定鸭疫里默氏杆菌中与卡那霉素抗性相关的基因,本研究采用构建Yb2株转座子随机突变库结合卡那霉素抗性平板来筛选相关抗性基因。先以Yb2株为受体菌,携带质粒pEP4351的大肠杆菌BW19851(pEP4351)为供体菌,构建转座子Tn4351随机突变库。用含红霉素和卡那菌素的TSA进行筛选,得到卡那霉素耐药性降低的突变株DK-2;然后采用基因组步移的方法鉴定表明,转座子插入基因组上编码核糖50S L27亚基基因;再将携带野生株Yb2的50S L27基因的大肠杆菌-鸭疫里默氏杆菌穿梭表达载体pRES-50S L27导入突变株DK-2,构建得到了回复菌株cDK-2。结果表明:50S L27基因缺失对其生长无明显的影响,而DK-2突变株对卡那霉素、新霉素和利福平的敏感性均增强。本研究为进一步解析鸭疫里默氏杆菌50S L27亚基的功能奠定了基础。In order to identify kanamycin resistance-related genes in Riemerella anatipestifer,random mutation library of Yb2 transposon was constructed and screened with kanamycin resistance TSA.A library of random Tn4351 transposon mutants was constructed using Yb2 strain as recipient and E.coli BW19851(pEP4351)as donor.The mutant DK-2 with decreased kanamycin resistance was screened by TSA containing erythromycin and kanamycin.The insertion site of Tn4351 transposon on the DK-2 genome was identified to be the gene encoding the 50S L27 subunit of ribosome.Subsequently,the shuttle expression vector pRES-50S-L27 carrying the 50S-L27 wild type Yb2 was introduced into the mutant DK-2 by bi-mating,and the complemented strain cDK-2 was then constructed.The analysis of biological characteristics showed that the insertion deletion of 50S L27 gene had no significant effect on its growth while the sensitivity of mutant DK-2 to kanamycin,neomycin and rifampicin increased.This study was helpful to elucidate the function of 50S L27 subunit ofR.anatipestifer.

关 键 词：鸭疫里默氏杆菌 卡那霉素抗性 转座子随机突变 核糖体50S L27亚基 回复株 

分 类 号：S852.612[农业科学—基础兽医学]