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为了制备抗华支睾吸虫基因工程抗体,本文从感染华支睾吸虫患者的淋巴细胞中提取总RNA,逆转录成cDNA。用相应的引物进行PCR,扩增出约700bp的重链Fd段和轻链κ、λ基因,经Xho I和Spe I,SacI和 Xba I双酶切后,分别和质粒载体pComb3连接,再经电穿孔转化大肠杆菌XL 1-blue菌株,将轻链和重链Fd基因先后克隆人pComb3中,成功地构建了抗华支睾吸虫Fab段抗体基因的表达载体,为进一步构建噬菌体抗体库奠定基础。
In order to prepare anti-Clonorchis sinensis genetically engineered antibodies, we extracted total RNA from lymphocytes infected with Clonorchis sinensis and reverse-transcribed them into cDNA. The corresponding primers were used to amplify the heavy chain Fd fragment of about 700 bp and the light chain κ and λ genes. After digested with Xho I, Spe I, SacI and Xba I, the plasmid was ligated with the plasmid vector pComb3, The E. coli XL 1-blue strain was transformed by electroporation. The light chain and heavy chain Fd genes were cloned into human pComb3. The expression vector of Fab fragment against Clonorchis sinensis was successfully constructed, which laid the foundation for the further construction of the phage antibody library .