根据NCBI上已公布的枯草芽孢杆菌BS168菌株的β-1,4-glucanase(eglS)基因以及苏云金芽孢杆菌的aiiA基因序列,设计引物并扩增得到eglS基因的启动子、信号肽序列和aiiA基因.构建重组表达载体Ps4-eaiiA,转化枯草芽孢杆菌BS168,得到枯草芽孢杆菌工程菌BS168/Ps4-eaiiA.该工程菌诱导发酵后经SDS-PAGE和Western blotting分析表明:aiiA基因实现了在枯草芽孢杆菌中的表达.对胡萝卜软腐欧文氏菌进行马铃薯的抗病性实验,结果表明aiiA基因在枯草芽孢杆菌中表达的AiiA蛋白对胡萝卜欧文氏杆菌表现出一定的抗病性.
Specific PCR primers were designed based on the β-1 ,4-glucanase (eglS) gene and aliA gene sequence reported in NCBI. The promoter and signal peptide of the eglS gene and aiiA gene were successfully amplified. Recombinant expression vector transformed Bacillus subtilis BS168, obtained engineering bacterium Ps4-eaiiA. SDS-PAGE and Western blotting suggested that Bacillus subtilis expressed the AiiA protein successfully in fermentation solution. Potato disease re- sistance experiment showed that AiiA protein expressed by Bacillus subtilis BS168/Ps4-eaiiA had strong antimicrobial activity for Erwinia carotovora.