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目的:从大肠癌病人淋巴结中扩增抗体Fab段重链(Fd)和轻链(k)基因。方法以一步法提取淋巴结总RNA,采用逆转录及PCR反应。反应产物以琼脂糖凝胶电泳观察。结果:扩增的Fd和k基因长度分别为850和700bP,相应肿瘤和正常切缘组织未见扩增。结论:Fab段重链(Fd)和连链(k)基因的成功扩增为大肠癌基因工程抗体的制备噬菌体抗体库的构建奠定丁基础。
Objective: To amplify the Fab heavy chain (Fd) and light chain (k) genes from the lymph nodes of colorectal cancer patients. Methods One-step method was used to extract lymph node total RNA and reverse transcription and PCR were used. The reaction product was observed by agarose gel electrophoresis. RESULTS: The lengths of the amplified Fd and k genes were 850 and 700 bP, respectively. No amplification was observed in the corresponding tumors and normal margin tissues. Conclusion: The successful amplification of the heavy chain (Fd) and chain (k) genes in the Fab segment lays a foundation for the construction of a phage antibody library for the preparation of colorectal cancer genetically engineered antibodies.