摘要
为探索人可溶性增殖诱导配体 (sAPRIL)在多种肿瘤细胞的增殖和存活以及促肿瘤形成中的作用 ,用RT PCR从扁桃体总RNA中扩增出人sAPRIL基因 .经克隆测序后进行同源性比较 ,证实所克隆的基因即为sAPRIL .将克隆载体经酶切并构建表达载体 ,在大肠杆菌中表达 ,表达量达4 3 6 % .纯化蛋白后进行3 H TdR参入实验 ,表明sAPRIL有明显促进肿瘤的形成及肿瘤细胞的增殖与存活的作用 .
To obtain human soluble a proliferation inducing ligand (APRIL) cDNA and stable expression in E.coli DH5α,the total RNA,extracted from human tonsil,was amplified into the sequence of cDNA encoding sAPRIL by RT PCR and cloned into vector pUC18 for sequencing. Next,the identified cDNA was cloned into the new type of prokaryotic expression vector pQE 80L. Then the combined plasmids was transmited into E.coli DH5α and the sAPRIL molecule was successfully expressed in E.coli DH5α.The activity of this protein was identified by 3H TdR and it can obviously stimulated the growth of tumor cells.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2005年第1期61-64,共4页
Chinese Journal of Biochemistry and Molecular Biology
关键词
增殖诱导配体
基因克隆
大肠杆菌表达
肿瘤细胞增殖
a proliferation-inducing ligand, gene clone, E.coli expression, tumor cell growth
