摘要
对一株致病性的非结核分支杆菌(NTM)即偶发分支杆菌(M.fortuitum)建立基因组文库,选用同尾酶Sau3 AⅠ和BamHⅠ分别酶切基因组DNA和载体pUC18,回收1 kb^7 kb大小范围的插入性DNA片段,与已经酶切去磷酸化的载体用T4连接酶连接,并转化入DH5α感受态细胞中,进行蓝白斑和氨苄抗性筛选,用PCR方法检测连接效率。结果得到阳性克隆4×104个,达到基因组文库要求,用真空冷冻干燥技术保存。该文库的建立为以后对该菌特殊基因方面的研究及基因结构和调控等研究奠定了良好基础,同时也为其他NTM的研究工作奠定了一定的技术和理论基础。
Construction of DNA Library of Mycobacterium fortuitum that is a Nontuberculous mycobacteria.The DNA was digested with Sau3AⅠ and the pUC18-vector was digested with BamHⅠ.1 kb^7 kb DNA fragments were recovered.The DNA fragments and pUC18-vector which was digested and removed phosphate groups were linked with T4 ligase.Using the color reation of α-complemnetation and resistance of ampicillin screened the linking production which had transformed into DH5α.Construction of the efficiency of linking was detected by polymerase chain reaction of positive strain.The number of positive bacterial recombinants obtained was 4×104.The construction of DNA library consistented with standards.We tried to conserve DNA library by using vacuum freeze-drying and this study was at stage of stability surveying.The DNA library provided help in studying and controlling of unusual gene of Mycobacterium fortuitum as well as gaving datas of technique and theory in the research on others nontuberculous mycobacteria（NTM）.
出处
《动物医学进展》
CSCD
北大核心
2011年第4期10-14,共5页
Progress In Veterinary Medicine
基金
国家科技支撑计划项目(2007BAD55B05)
