地衣芽孢杆菌16S-ITS标记及其特异性分析

摘要

[Objective] This study aimed to establish molecular identification methods for Bacillus licheniformis. [Method] Based on clone sequencing and difference analysis for 16S and ITS sequences of B. licheniformis TS-01, specific primers were designed using region sequences as the targets used for amplifying all test strains. [Result] The specific primers of B. licheniformis were designed from the ITS and 16S rDNA regions. The optimal annealing temperature of the specific primers for PCR was 67.2 ℃ with 24 cycles. A 905 bp marker fragment was amplified for B. licheniformis TS-01, while all other test strains showed negative results. This indicated that a specific 16S-ITS marker was obtained, which accurately identified the strain at the species level. [Conclusion] This molecular identification method for B. licheniformis TS-01 has laid the foundation for molecular diagnosis of B. licheniformis.%[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16Sr DNA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905bp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。