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目的 获得人甘露聚糖结合凝集素相关丝氨酸蛋白酶－２（ＭＡＳＰ－２）编码区基因。方法采用 ＲＴ－巢式 ＰＣＲ技术从 人胎肝组织总ＲＮＡ扩增目的cＤＮＡ片段，克隆人ｐＧＥＭ－Ｔ载体，酶切图谱分析和测序鉴定。结果获得了编码合信号 顺序的ＭＡＳＰ－２肽链全长cＤＮＡ，将其与pＧＥＭ－Ｔ载体连接，转化大肠杆菌ＴＧ１，建立了ＭＡＳＰ－２的重组克隆。酶切图 谱与微机分析结果无异；序列分析表明，与报道的人 ＭＡＳＰ－２ｃＤＮＡ序列一致。结论成功克隆了人 ＭＡＳＰ－２基因，为深 入研究补体凝集素途径的激活机制和甘露聚糖结合凝集素基因突变引起免疫缺损的发病机制奠定了基础。
Objective To obtain the coding region of human mannan-binding lectin-associated serine protease-2 (MASP-2). Methods The target cDNA fragment was amplified from human fetal liver by RT-nested PCR and cloned into pGEM-T vector. The restriction enzyme digestion analysis and sequencing were performed. Results The full-length cDNA of MASP-2 peptide was obtained. The full-length cDNA of MASP-2 was cloned into pGEM-T vector and transformed into E.coli TG1. The recombinant MASP-2 was cloned. The results of the enzyme digestion map were the same as those of the computer analysis; the sequence analysis showed that it was consistent with the reported human MASP-2 cDNA sequence. Conclusion The human MASP-2 gene was successfully cloned, which laid the foundation for further study of the activation mechanism of complement lectin pathway and the pathogenesis of immunodeficiency induced by mannan-binding lectin gene mutation.