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目的:为了获得人CD59cDNA完整序列,以建立表达人CD59分子的小鼠T细胞模型,更深入地探讨补体MAC及CD59分子与T细胞活化的关系。方法:通过设计和合成引物、提取细胞总RNA进行逆转录反应,经PCR扩增目的片段,并将其克隆于pUC18及pUC19质粒上,测定其DNA序列。结果:从Jurkat细胞总RNA中扩增得到396bp片段,序列测定证实该片段为包括25a信号肽在内的全部CD59编码序列。结论:表明CD59cDNA已成功地得到克隆。
OBJECTIVE: In order to obtain the complete sequence of human CD59 cDNA to establish mouse T cell model expressing human CD59 molecule, we further investigate the relationship between complement MAC and CD59 molecules and T cell activation. Methods: Total RNA was extracted from the cells for reverse transcription reaction by designing and synthesizing primers. The target fragment was amplified by PCR and cloned into pUC18 and pUC19 plasmids. The DNA sequence was determined. Results: A 396 bp fragment was amplified from total RNA of Jurkat cells. Sequence analysis confirmed that the fragment was all CD59 coding sequence including 25a signal peptide. Conclusion: This demonstrates that CD59 cDNA has been successfully cloned.