CAJ | 학술논문

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,克隆入pGEM-T载体,酶切图谱分析和测序鉴定。结果获得了编码含信号顺序的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-2 cDNA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。
목적획득인감로취당결합응집소상관사안산단백매-2(MASP-2)편마구기인。방법채용RT-소식PCR기술종인태간조직총RNA확증목적cDNA편단,극륭입pGEM-T재체,매절도보분석화측서감정。결과획득료편마함신호순서적MASP-2태련전장cDNA,장기여pGEM-T재체련접,전화대장간균TG1,건립료MASP-2적중조극륭。매절도보여미궤분석결과무이;서렬분석표명,여보도적인MASP-2 cDNA서렬일치。결론성공극륭료인MASP-2기인,위심입연구보체응집소도경적격활궤제화감로취당결합응집소기인돌변인기면역결손적발병궤제전정료기출。
Objective To obtain the gene fragment encoding human mannan-binding lectin (MBL)-associated serine protease-2 (MASP-2). Methods The target cDNA fragment was amplified from the total RNA extracted from human fetal liver tissue by RT-nested PCR, cloned into pGEM-T vector and identified by restriction analysis and sequencing. Results The cDNA fragment encoding MASP-2 polypeptide with the signal sequence was isolated, linked with pGEM-T vector and transformed into E.coli TG1. The restriction maps of the selected clones were consistent with those generated by computer analyses. The result of sequencing of one selected clone showed that its sequence was identical with that of reported human MASP-2 cDNA. Conclusion The human MASP-2 gene was successfully cloned, which prepares the ground for studying the molecular mecha- nisms by which MBL activates the complement lectin pathway and the mutations of MBL gene cause immunodeficiency.