摘要
利用从Bacillus pumilus 289中分离出的隐秘质粒pNK289(7.2kb)的复制起始调控区及启动区DNA片段和质粒pPL 601上的氯霉素乙酰基转移酶结构基因(cat-86),构建了能在B.subtilis和B. pumilus中稳定传代的质粒pNQ216(4.1kb)和pNQ402(2.8kb)。在非诱导条件下,其在含Cm的LB平皿上的外显率都比pPL600高约30％,可以用作芽孢杆菌基因克隆的质粒载体。
Two new plasmids pNQ216 (4.1kp) and pNQ402 (2.8kb) were constructed by combining the replication origin of the plasmid pNK289,a cryptic plasmid resides in B.pumilus 289,and the cat-86 gene from plasmid pPL601.These two plasmids can be maintained steadily in both B.subtilis and B.pumilus.The penelrance of these plasmids on LB medium with Cm (20μg/ml) is 30% higher than that of pPL600.Thus both of these plasmids can be used as new cloning vectors in Bacillus.
关键词
芽孢杆菌
隐秘质粒
载体构建
Cryptic plasmid,Bacillus pumilus,Bacillus subtilis,Vector construction
