摘要 目的 :对独特型抗体可变区基因进行序列分析,为其基因工程抗体的构建打下基础。方法 :采用TRIzolR试剂从分泌抗独特型抗体SM6杂交瘤细胞株中提取mRNA,以特异性引物采用PCR技术扩增并克隆出独特型单抗SM6重、轻链可变区(VH、VL)基因。将VH和VL基因分别克隆到pGEM-T载体中。采用链末端终止法荧光染色测定VH和VL基因序列,应用DNASIS7分析软件并与美国国立卫生研究院基因库比较分析。结果 :5个克隆的重链基因序列一致,3个克隆的轻链基因中有2个克隆基因一致。结论 :VH和VL基因分别属Q52、JH3和VK19、JK2。

DNA sequencing of variable regionsof anti-idiotypic monoclonal antibody SM6 to B-CLL SuNa, Yang Jing, Wang Xiaolin, et al. Department of Immunology, Tongji Medical University,Wuhan 430030

    Abstract Objective:Toanalyze the variable region sequences of an anti-idiotypic monoclonal antibody to humanchronic B cell leukemia (B-CLL). Methods:The variable region sequences of light and heavychains (VL、VH) of anti-idiotypic McAb SM6 to B-CLLwere analyzed by their cDNA cloning and sequencing. Total RNA was extracted from ahybridoma cell line producing SM6. The VH and VL genes were amplified by RT-PCR withspecific primers. The PCR products were cloned into pGEM-T vectors, then transfected intoJM109. Results and Conclusion: It was confirmed by DNASIS7 sequence analysis that thefull-length and potentially functional genes were successfully cloned. The VH geneexpressed in SM6 belongs to Q52, the JH region using the JH3. The VL gene belongs to VK19,the JK region using JK2.

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