作　　者： ; ; ; ; ; ; ;

机构地区： 中山大学生命科学学院有害生物控制与资源利用国家重点实验室

出　　处： 《中山大学学报（自然科学版）》 2006年第4期79-82,共4页

摘　　要： 根据苏云金杆菌(Bacillus thuringiensis,B.t)溶原性噬菌体G IL01(GenBank Accession Number:AJ536703)的同源序列设计一对引物,用35株B.t标准菌株和生产菌株MZ1(血清型H3 a3b)的过夜培养物作模板,结果有16株菌株包括MZ1能扩增出大小约为750 bp的预期产物,推测这16株菌为溶原菌。MZ1菌株的过夜培养液与指示菌ZK1(血清型H25)混匀并在双层琼脂上培养12 h后出现了滴度为2×107pfu的噬菌斑,从而证明生产菌株MZ1确为溶原菌;又以G IL01同源性较高的其它四个基因设计引物,分别以MZ1的培养物及其溶原性噬菌体基因组为模板进行PCR扩增,结果两者得到几乎一致的带型,进一步证明了生产菌株MZ1为溶原菌,说明利用PCR法检测鉴定苏云金杆菌的溶原性是可行的。比较指示菌方法、直接电镜法和DNA鉴定法,PCR法快速简便,对B.t生产上快速检测其溶原性噬菌体有实用意义。 According to one conserved sequences of phage GIL01 from Bacillus thuringiensis （B. t）, a pair of primers was designed. With overnight cultivations of 35 B. t standard strains and one fermentative strain MZ1 as templates, PCR results showed that 16 strains, which may be lysogenic strains, produced the product with the size about 750 bp as anticipated. Among them strain MZ1 had been confirmed a lysogenic strain by indicator strain ZK1 after producing phage plaques induced by Mitomycin C. Two templates of overnight cultivation and its DNA from strain MZ1 were then amplified in the same time with five pairs of primers designed from the same phage, and the same products were obtained, suggesting that strain MZ1 was also confirmed a lysogenic strain by PCR method. Compared with other methods to detect B. t lysogeny, PCR is simpler and less time comsuming. In particular, it is a practical method for the quick detection in B. t fermentation.

关 键 词： 苏云金杆菌 溶原性 噬菌体

领　　域： [生物学] [生物学]