抗菌肽基因misgurin同向串连表达载体构建新策略

A new strategy in the construction of expression vector with multi-copy misgurin gene tandem in the same direction

目的是将抗菌肽基因misgurin以同向串连方式连接成多拷贝基因 ,并克隆到甲醇酵母表达载体上。在设计和连接中采用新策略 ,运用常规分子克隆操作进行表达载体构建 ,结果表明 ,该策略能方便高效地进行基因多拷贝同向串连表达载体的构建。

A new strategy was adopted in the procedure of designing and ligating In the synthesis of misgurin gene and adapters,the codon Met(ATG/TAC) was added into the oligoDNA as a cohesive end,which would make the genes tandem in the same direction In the procedure of ligating,misgurin gene was ligated with two adapters respectively firstly,the gene was added every six hours instead of all in one time They were mixed until these productions with likely length formed At the same time,PEG8000 at the concentration of 15% was added in the ligating reaction system The result showed that multi-copy gene expression vector can be constructed conveniently and efficiently with the new strategy