地衣芽孢杆菌16S-ITS标记及其特异性分析  

Analysis on 16S-ITS Marker from Bacillus licheniformis and Its Specificity

作　　者：陈冲[1] 佟建明[2] 张潞生[3] 

机构地区：[1]山西农业大学信息学院,山西太谷030801 [2]中国农业科学院北京畜牧兽医研究所,北京100094 [3]中国农业大学农学与生物技术学院,北京100094

出　　处：《安徽农业科学》2011年第14期8206-8207,共2页Journal of Anhui Agricultural Sciences

摘　　要：[目的]建立地衣芽孢杆菌的分子鉴定方法。[方法]通过对地衣芽孢杆菌TS-01的16S和ITS序列进行克隆测序和差异性分析,以该区序列为靶序列设计地衣芽孢杆菌TS-01的特异性引物,并用该引物扩增全部受试菌种。[结果]从ITS和16S rDNA区间设计了地衣芽孢杆菌种特异性引物,特异性引物PCR的最佳退火温度为67.2℃,循环数为24个;地衣芽孢杆菌TS-01可扩增出1条905 bp的标记片段,其他受试菌株均为阴性,从而证明试验得到了在种水平上对该菌种进行准确鉴定的特异16S-ITS标记。[结论]地衣芽孢杆菌TS-01分子鉴定方法的建立,为地衣芽孢杆菌的分子诊断奠定了基础。[Objective]The study aimed to establish the molecular identification methods of Bacillus licheniformis.[Method]Through the cloning sequencing test and the difference analysis on 16S and ITS sequences of B.licheniformis TS-01,the specific primer of B.licheniformis TS-01 was designed with the sequence of the above regions as the target sequence and the whole tested strains were amplified by using this specific primer.[Result]The specific primers of B.licheniformis were designed resp.from the DNA section of 16S and from that of ITS and the optimum annealing temperature for the specific primer PCR was 67.2 ℃ with the cycle times of 24.B.licheniformis TS-01 could amplify a marker fragment of 905 bp and other tested strains were all negative,thus proving that this test obtained the specific 16S-ITS marker that could accurately identify this strain at the level of the specie.[Conclusion]Establishment on the molecular identification methods of B.licheniformis TS-01 laid the foundation for molecular diagnosis of B.licheniformis.

关 键 词：地衣芽孢杆菌 特异引物 16S ITS 

分 类 号：S816.3[农业科学—饲料科学]