CAJ | 학술논문

细胞核因子κB受体活化因子配体（Receptor activator of the NF -κB ligand ， RANKL）是TNF超家族的重要成员之一，属于Ⅱ型跨膜蛋白，通过与其受体核因子κB受体活化因子（RANK）构建的信号通路参与乳腺癌的发生、肿瘤骨转移、骨质疏松、关节炎等病理过程。人RANKL胞外结构域基因片段与原核表达载体pET-21b融合并转化至表达菌Rosetta，并对其表达条件进行了优化。通过对诱导时机，诱导温度，异丙基-β-D-硫代吡喃半乳糖苷（IPTG）浓度，诱导时间及甘油浓度的条件优化表明，当IPTG的浓度为0．2 mmol/mL，20℃振荡诱导培养6 h 时，甘油浓度为4％时，可在上清中获得高效表达的pET-21b-RANKL融合蛋白，为该蛋白的进一步纯化及结构与功能研究打下了良好的基础。
세포핵인자κB수체활화인자배체（Receptor activator of the NF -κB ligand ， RANKL）시TNF초가족적중요성원지일，속우Ⅱ형과막단백，통과여기수체핵인자κB수체활화인자（RANK）구건적신호통로삼여유선암적발생、종류골전이、골질소송、관절염등병리과정。인RANKL포외결구역기인편단여원핵표체재체pET-21b융합병전화지표체균Rosetta，병대기표체조건진행료우화。통과대유도시궤，유도온도，이병기-β-D-류대필남반유당감（IPTG）농도，유도시간급감유농도적조건우화표명，당IPTG적농도위0．2 mmol/mL，20℃진탕유도배양6 h 시，감유농도위4％시，가재상청중획득고효표체적pET-21b-RANKL융합단백，위해단백적진일보순화급결구여공능연구타하료량호적기출。
The receptor activator of nuclear factor -kappa B ligand (RANKL) is an important member of the TNF superfami-ly, which is a type II transmembrane protein , and its cognate ligand, RANK (receptor activator of nuclear factor -kappa B) built the signaling pathway in the occurrence of breast cancer , bone metastases, osteoporosis, arthritis and other pathological processes.In this article, the extracellular domain of human RANKL gene were cloned into prokaryotic expression vector pET-21b and transformed into the expression bacteria E .coli Rosetta.We successfully constructed recombinant human RANKL gene Rosetta/pET-21b-RANKL.And it was induced to express the target protein .We optimized its expression conditions . When the IPTG concentration was 0.2 mmol/mL, the induced timing OD600 for 0.6, oscillation induced culture for 6 h in 20℃and glycerol concentration for 4%, we can efficiently expressed the extracellular domain of human RANKL fusion protein (pET-21b-RANKL) in the supernatant.It provided a foundation for further purification of the protein and researched its structure function.