芽孢杆菌TS02特异RAPD标记的SCAR标记转化和验证

Transformation and Identification of SCAR Marker from Specific RAPD Marker of Bacillus TS02

摘要:

利用自主分离的蜡状芽孢杆菌菌株TS02, 采用RAPD方法对TS02及其同源性相近的5株芽孢杆菌(地衣芽孢杆菌、枯草芽孢杆菌、凝结芽孢杆菌、巨大芽孢杆菌、短小芽孢杆菌) 进行了RAPD条带特异性分析, 从TS02基因组中筛选获得了一个533 bp的特异RAPD标记TSR1。TSR1克隆、测序后, 根据其序列设计出一对特异引物P1/P2进行扩增, 结果只在TS02中扩增得到目的片段, 而其余对照菌株扩增为阴性, 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SCAR标记。

Abstract:

Bacillus cereus TS02 is a functional strain separated by our laboratory. RAPD was used to analyse the specificity of TS02 in contrast to other five Bacillus bacteria (Bacillus licheniformis, Bacillus subtilis, Bacillus coagulans, Bacillus megaterium and Bacillus pumilus) that are highly homologic to TS02 according to the conventional taxonomy. A specific marker TSR1 (533 bp) was obtained, cloned and sequenced, a specific primer pair P1/P2 was designed based on the TSR1 sequence. P1/P2 can specifically amplify the objective band which is only in TS02 and not found in other control strains. The results showed that the specific RAPD marker had been transformed to the steady SCAR marker, and the specific SCAR marker can correctly detect the Bacillus TS02 in specie’s lever.