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用RTPCR方法从人单核THP1细胞系克隆人41BBL胞外区基因,将其重组到pAYZ表达载体中,构建成人41BBL胞外区基因表达载体。将该载体转化大肠杆菌16C9,获得稳定表达,表达产物主要以可溶性状态存在;SDSPAGE和Westernblot分析显示,其分子量约为22kD,与预期结果一致。这是首次在大肠杆菌中获得41BBL胞外区可溶性表达。生物学活性检测显示41BBL对于维持T淋巴细胞系因子释放非常有益,同时PI单染表明它能抑制Jurkat细胞的凋亡。这将在抗肿瘤免疫治疗中具有潜在应用前景。
The human 41BBL extracellular region gene was cloned from human mononuclear THP1 cell line by RTPCR method and recombined into pAYZ expression vector to construct the human 41BBL extracellular region gene expression vector. The vector was transformed into Escherichia coli 16C9 to obtain a stable expression, the expression product mainly exists in a soluble state; SDSPAGE and Western blot analysis showed that the molecular weight of about 22kD, with the expected results. This is the first time that 41BBL extracellular domain soluble expression was obtained in E. coli. The biological activity tests showed that 41BBL was very helpful for the maintenance of T lymphocyte factor release, while the PI single staining indicated that it could inhibit the apoptosis of Jurkat cells. This will have potential applications in antitumor immunotherapy.