抗猪囊尾蚴头节特异性单链抗体的构建及克隆

Constructing and cloning of single-chain antibody against Cysticercus Cellulosae scolex antigen

首先对猪囊尾蚴头节特异性单链抗体重链可变区基因(Vh)和轻链可变区基因(Vl)进行酶切鉴定,PCR鉴定以及测序分析,然后采用重叠延伸PCR(SOE PCR)技术,将重链可变区基因和轻链可变区基因,通过linker链连接成Vh-linker-Vl单链抗体基因(ScFv)。将此基因与pMD18-T载体连接,转化感受态大肠杆菌J M109,从而进一步克隆并对重组载体pMD-ScFv进行鉴定。结果:成功构建并克隆了抗猪囊尾蚴头节单链抗体,为重组免疫毒素ScFv-PE40的构建?表达及活性测定奠定了良好的基础。

First enzyme-cutting,PCR analysis and sequence determination for variable region gene of heavy chain（Vh） and light（Vl） of single-chain antibody against Cysticercus Cellulosae Scolex.Then the single-chain antibody gene（ScFv） was constructed by Vh,linker and Vl by gene splicing by over lap extension PCR（SOE PCR）.The amplified products were linked to vector pMD-18T to carry out its sequence determination,then compared this sequence with that of strain isolated abroad.The vector was proved through the identification of enzyme-cutting and PCR analysis.The result indicated that it is successful to construct and clone the single-chain antibody against Cysticercus Cellulosae scolex antigen.The product could serve as a basis for constructing,expressing and activity detecting of recombinant immunotoxin ScFv-PE40.