双价抗菌肽基因表达载体的构建

CONSTRUCTION OF BIVALENT CECROPIN GENE EXPRESSION VECTOR

将人工合成的抗菌肽Ｄ基因和抗菌肽Ｂ基因克隆到同一植物表达载体ｐＢⅠ１２１，且各自具有其３５ｓ启动子和Ｎｏｓ终止子。先将抗菌肽Ｄ基因及其启动子和终止子克隆到ｐＢ１２１ＨｉｎｄⅢ位点获得ｐＥＣＶⅠ重组子。然后将抗菌肽Ｂ基因克隆到ｐＥＣＶⅠ的ＢａｍＨⅠ位点，经Ａｇａｒｏｓｅｇｅｌ检测和酶切鉴定获得具有抗菌肽Ｄ，抗菌肽Ｂ基因的双价基因表达载体ｐＥＣＶⅡ。

The present study tried to construct a bivalent Cecropin gene plantexpression vector. Cecropin D gene and Cecropin B gene synthesized werecloned into the san1e plant expression vector called pBI121.Cecropin D geneDNA fragments with CaMV 35s promotor and Nos terminor were cloned intoOBl121 HindIII site to obtain the recombinant pCEVI by screening,Cecropin Bgene DNA fragments were then cloned into pECVI BamHI site,and a bivalentplant expression vector caIled pECVI was obtained through agarose gel elec-trophoresis and enzyme digestion. This vector contains Cecropin D geneand Cecropin B gene each having a CaMV 35s promotor and a Nos terminor.