摘要
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:CD59,with multiple functions,is a widely expressed cell surface glycosyl phosphatidyl inositol(GPI) anchored glycoprotein.It acts as an inhibitor of the assembly of the membrane attack complex of autologous complement,binds to CD2,and also transduces activation signals in T cells.The purpose of this study is to obtain the cDNA sequence encoding humam CD59.Methods:The primers are designed based on the known CD59 cDNA sequence.Total RNA was isolated from cultured Jurkat cells,and then RT PCR was performed.The fragment was cloned into pUC18 and pUC19 plasmids,and further sequenced by sanger's dideoxy mediated chain termination.Results:A 396 bp DNA fragment was amplified by RT PCR method from Jurkat cells.This cDNA fragment includes 384 bp open reading fragment and it's sequence is identical to the published sequence encoding human CD59.Conclusion:CD59 cDNA has been cloned successfully.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
1999年第3期97-99,共3页
Chinese Journal of Immunology
基金
国家自然科学基金
