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抗慢性淋巴细胞白血病独特型单抗SM6可变区基因序列分析
DNA sequencing of variable regions of anti-idiotypic monoclonal antibody SM6 to BCLL
【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pGEMT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。
【Abstract】 Objective:To analyze the variable region sequences of an antiidiotypic monoclonal antibody to human chronic B cell leukemia (BCLL). Methods:The variable region sequences of light and heavy chains (VL、VH) of antiidiotypic McAb SM6 to BCLL were analyzed by their cDNA cloning and sequencing. Total RNA was extracted from a hybridoma cell line producing SM6. The VH and VL genes were amplified by RTPCR with specific primers. The PCR products were cloned into pGEMT vectors, then transfected into JM109. Results and Conclusion: It was confirmed by DNASIS7 sequence analysis that the fulllength and potentially functional genes were successfully cloned. The VH gene expressed in SM6 belongs to Q52, the JH region using the JH3. The VL gene belongs to VK19, the JK region using JK2.
【Key words】 Leukemia Antibody; anti-idiotype Sequence analysis Gene cloning;
- 【文献出处】 中华血液学杂志 ,CHINESE JOURNAL OF HEMATOLOGY , 编辑部邮箱 ,1998年07期
- 【分类号】R733.7
- 【被引频次】1
- 【下载频次】14