[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.
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