CAJ | 학술논문

Mimecan/osteoglycin是一种分泌型蛋白质,目前其功能尚不明确.从人垂体cDNA中克隆到OIF基因并构建成重组表达质粒pGEX-5X-2/mimecan,将此重组表达质粒转化大肠杆菌BL21(DE3)后用IPTG诱导,成功表达了一种分子量约为38 kD融合蛋白,约占菌体总蛋白的30%～40%.此融合蛋白经纯化分离后免疫新西兰大白兔以制备多克隆抗体.用Western印迹法检测兔的抗血清.结果显示,该多克隆抗体有较好的针对mimecan蛋白的专一性并且效价较高,可用于对mimecan的功能研究.用此多克隆抗体检测到在某些种类的人垂体瘤组织中mimecan表达极高,提示可能存在新的垂体瘤类型.
Mimecan/osteoglycin시일충분비형단백질,목전기공능상불명학.종인수체cDNA중극륭도OIF기인병구건성중조표체질립pGEX-5X-2/mimecan,장차중조표체질립전화대장간균BL21(DE3)후용IPTG유도,성공표체료일충분자량약위38 kD융합단백,약점균체총단백적30%～40%.차융합단백경순화분리후면역신서란대백토이제비다극륭항체.용Western인적법검측토적항혈청.결과현시,해다극륭항체유교호적침대mimecan단백적전일성병차효개교고,가용우대mimecan적공능연구.용차다극륭항체검측도재모사충류적인수체류조직중mimecan표체겁고,제시가능존재신적수체류류형.
Mimecan belongs to a family of leucine-rich proteoglycans that are secreted into the extracellular matrix. In order to investigate the function of mimecan, the coding region of mimecan was amplifed from a human pituitary cDNA by PCR and the recombinant prokaryotic expression vector pGEX-M was constructed. The vactor was transformed into E.coli BL21(DE3)and the GST-M fusion protein of 38 Kd was ecpressed in the bacteria under induction of IPTG. After purification, the fusion protein was infucted into New Zealand rabbits to prepare polyclonal antibody. The antibody was tested by Western blotting for their specificity and sensitivity. Using the antibody it was found the mimecan was expressed highly in certain types of human pituitary tumor tissues. These results make it possible for studying the biological function of mimecan.