中华蜜蜂Apisimin基因的克隆及在大肠杆菌内的表达  

Molecular Cloning and Expression in E.coli of Apisimin cDNA from Apis cerana cerana

作　　者：杜宏沪[1,2] 苏松坤[1] 梁勤[2] 陶挺[1] 

机构地区：[1]浙江大学动物科学学院,浙江杭州310029 [2]福建农林大学蜂学学院,福建福州350002

出　　处：《蜜蜂杂志》2007年第S1期43-46,共4页Journal of Bee

摘　　要：构建了中华蜜蜂8日龄工蜂头部cDNA文库,筛选获得Apisimin基因。该基因cDNA全长407 bp,含一个243 bp的开放阅读框(ORF),编码78个氨基酸的多肽,其N端含有一个由24个氨基酸残基组成的信号肽,预测成熟肽的分子量为5.9 kDa。中蜂Apisimin基因与西方蜜蜂和印度蜜蜂Apisimin的相似性分别为93%和100%。采用BamH I和Xho I双酶切位点将克隆获得的Apisimin基因连接到pET30a表达载体上,筛选出含有目的基因的阳性菌株,经IPTG诱导表达,收集菌体进行SDS-PAGE和Western印迹鉴定。为进一步研究Apisimin的结构和功能提供依据。A cDNA library was constructed from 8-day worker heads of Apis cerana cerana and the apisimin gene was screened from it.The cDNA of apisimin is 407bp and contains an open reading frame (ORF) of 237bp,encoding 78 amino acid residue peptide with a signal peptide of 24 amino acid in the N-terminal.The molecular weight of mature apisimin is 5.9 kDa.The similarity of Apis cerana cerana apisimin was 93% and 100% comparing with Apis mellifera and Apis cerana india,respectively.The cloned apisimin gene was linked into the pET30a expression vector using both BamH I and Xho I enzyme digestions.The positive E.coli BL21 (DE3) including Apisimin was screened and induced with IPTG to produce the peptide of Apisimin.The goal bacteria were collected used to SDS-PAGE analysis and western blot identification.Thus,our researches provide good conditions for further studies on the structure and function of Apisimin peptide.

关 键 词：中华蜜蜂 Apisimin 分子克隆 原核表达 

分 类 号：S891[农业科学—特种经济动物饲养]