CAJ | 학술논문

目的：探讨Matriptase-2重组蛋白的表达及多克隆抗体制备。方法 Matriptase-2胞外区近跨膜段（Glu81-Gly228）克隆入pMAL-C2X，重组质粒转化BL21(DE3)大肠杆菌，经IPTG诱导表达，提取细菌蛋白后用直链淀粉树脂纯化柱进行纯化。将纯化后的融合蛋白免疫新西兰大白兔，制备Matriptase-2抗血清；免疫血清用protein G亲和层析柱进行纯化，获得多克隆抗体。结果成功表达了Matriptase-2重组融合蛋白，获得了高效价的抗血清，Western blot结果显示具有抗原特异性识别。结论成功制备了高特异性、高效价的抗人Matriptase-2多克隆抗体，为今后深入研究Matriptase-2的生物学功能提供了有用的实验工具。
목적：탐토Matriptase-2중조단백적표체급다극륭항체제비。방법 Matriptase-2포외구근과막단（Glu81-Gly228）극륭입pMAL-C2X，중조질립전화BL21(DE3)대장간균，경IPTG유도표체，제취세균단백후용직련정분수지순화주진행순화。장순화후적융합단백면역신서란대백토，제비Matriptase-2항혈청；면역혈청용protein G친화층석주진행순화，획득다극륭항체。결과성공표체료Matriptase-2중조융합단백，획득료고효개적항혈청，Western blot결과현시구유항원특이성식별。결론성공제비료고특이성、고효개적항인Matriptase-2다극륭항체，위금후심입연구Matriptase-2적생물학공능제공료유용적실험공구。
Objective To investigate the expression of Matriptase-2 recombinant protein inE.coli and the generation of its polyclonal antibody.Methods Extracellular region (Glu81-Gly228) proximal to transmembrane domain of Matriptase-2 was cloned into pMAL-C2X. The recombinant plasmid was transformed to BL21 (DE3) bacteria. After IPTG induction, the total protein was extracted and the recombinant Matriptase-2 protein was purifi ed with amylose resin. The Matriptase-2 recombinant protein was used to immunize New Zealand Rabbits to prepare serum with anti-Matriptase-2 polyclonal antibody. The antibody was purifi ed from serum by protein G sepharose 4B.Results Recombinant Matriptase-2 protein was successfully expressed inE.coliand anti-serum with high titer was obtained. Western blotting result showed that the antibody specifi cally recognized Matriptase-2. Conclusion It is concluded that the anti-Matriptase-2 polyclonal antibody with high titer and specificity is successfully generated. The antibody provides an useful experimental tool to investigate the biological function of Matriptase-2.