家蚕抗菌肽基因Cecropin-D的克隆及其真核表达载体的构建  

Cloning and Construction of the Expression Vector of Antimicrobial Peptide Cecropin D gene from Bombyx mori

作　　者：章玉萍[1] 范涛[1] 马忠友[2] 代君君[1] 王储言 李瑞雪[1] 吴传华[1] 田善富[1] 肖林珍[1] 

机构地区：[1]安徽省农业科学院蚕桑研究所,合肥230061 [2]安徽科技学院,安徽凤阳233100

出　　处：《北方蚕业》2010年第4期18-21,共4页North Sericulture

摘　　要：从家蚕中提取总RNA,采用反转录聚合酶链式反应(RT-PCR)方法,获得了家蚕Cecropin-D基因编码区的cDNA,扩增出家蚕抗茵肽Cecropin-D成熟肽基因片段,重组克隆入pMD18-Tvector载体,经DNA测序,该基因为186bp,编码62个氨基酸。用限制性内切酶切下目的基因,插入毕赤酵母表达载体pPIC9K中,构建真核表达栽体pPlC9K-CD。本实验的研究为获得超量表达的高活性表达抗菌肽Cecropin-D以及为研制具有抗菌活性的新型基因药物奠定了基础。The total RNA was extracted from Bombyx mori,Cecropin-D gene cDNA was obtained by reverse transcription polymerase chain reaction（RT-PCR）,and then gene of the mature peptide of Cecropin-D was amplified and cloned into pMD18-T vector.The sequence analysis showed that the cloned DNA had a length of 192bp,encoding a protein of 64 amino acids.After digested by both EcoRI and NotI. DNA fragment was linked with vector plasmid pPIC9K by T4 DNA ligase. The construction of expression vector pPIC9-CD may lead to obtaining large Cecropin D,thus providing basis for further research on developing new gene drug of antibacterial activity.

关 键 词：家蚕 抗菌肽 Cecropin-D 克隆 表达栽体pPIC9K 

分 类 号：S881[农业科学—特种经济动物饲养]