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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 (κ)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κ基因长度分别为 70 0和 650bp ,相应肿瘤和正常切缘组织未见扩增。结论 Fab段重链 (Fd)和轻链 (κ)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了基础。
Objective To amplify the Fab heavy chain (Fd) and light chain (kappa) genes from the lymph nodes of colorectal cancer patients. Methods One-step lymph node total RNA was extracted and reverse transcription and PCR reactions were used. The reaction product was observed by agarose gel electrophoresis. Results The lengths of the amplified Fd and κ genes were 70 0 and 650 bp, respectively. No amplification was observed in the corresponding tumors and normal margin tissues. Conclusion The successful amplification of the Fab heavy chain (Fd) and light chain (kappa) genes laid the foundation for the construction of a phage antibody library for the preparation of colorectal cancer genetically engineered antibodies.