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目的：从一株抗结肠癌细胞系ＬＳ１７４Ｔ的单抗细胞株ＣＬ－４中克隆出抗结肠癌抗体重链可变区和κ轻链基因。方法：用一步法提取总ＲＮＡ，逆转录形成ｃＤＮＡ，设计并合成一套扩增小鼠抗体重链可变区和完整κ轻链的通用引物，通过ＰＣＲ扩增基因，分别克隆入ｐＵＣ１９，作核苷酸序列分析。结果：用重链引物扩增获得长约３６０ｂｐ的片段；用轻链引物扩增获得约６４０ｂｐ的片段，序列测定获得它们的ＤＮＡ序列。结论：克隆的重链可变区基因长度为３６０ｂｐ，属于小鼠重链可变区Ｉ（Ｂ）亚族；κ轻链长度为６４２ｂｐ，其中可变区为３２１ｂｐ，属于小鼠κ轻链Ⅴ亚族。
OBJECTIVE: To clone the anti-colon cancer antibody heavy chain variable region and κ light chain gene from a monoclonal antibody cell line CL-4 against colon cancer cell line LS174T. Methods: One-step total RNA was extracted and reverse transcribed to form cDNA. A set of universal primers for amplifying mouse antibody heavy chain variable region and complete κ light chain was designed and synthesized. The gene was amplified by PCR and cloned into pUC19. Nucleotide sequence analysis. RESULTS: A fragment of about 360 bp in length was amplified using heavy chain primers; fragments of about 640 bp were amplified using light chain primers, and their DNA sequence was determined by sequencing. Conclusion: The cloned heavy chain variable region gene has a length of 360bp and belongs to the mouse heavy chain variable region I (B) subfamily. The kappa light chain has a length of 642bp, of which the variable region is 321bp and belongs to the mouse κ light chain. Ethnic family.