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用套式PCR从人胎脑cDNA文库中克隆了B淋巴细胞刺激因子C端肽 (C terminalpeptideofBlymphocytestimulator,C BLyS)的cDNA。在大肠杆菌BL2 1CodonPlus (DE3)RIL中以包含体形式表达了C BLyS。对包含体的复性条件进行了摸索 ,建立了C BLyS的透析复性与纯化方法。经复性和纯化的C BLyS可结合其受体B细胞成熟抗原 (Bcellmaturationantigen ,BCMA) 人IgG1Fc融合蛋白 ,刺激体外培养的小鼠脾脏细胞增殖 ,并且可明显增强小鼠对溶菌酶的免疫应答水平。
CDNA of C terminal peptide of Baltimocytestimulator (C BLyS) was cloned from human fetal brain cDNA library by nested PCR. C BLyS was expressed as an inclusion body in E. coli BL2 1CodonPlus (DE3) RIL. The refolding conditions of the inclusion bodies were explored, and the dialysis refolding and purification methods of C BLyS were established. Refolded and purified C BLyS can bind to its receptor B cell maturation antigen (BCMA) human IgG1 Fc fusion protein to stimulate the proliferation of mouse spleen cells cultured in vitro, and can significantly enhance the mouse immune response to lysozyme level .