摘要
目的克隆、表达抑癌基因NDRG4,纯化后考察其抑瘤活性。方法将NDRG4基因克隆进PQE30/M15表达载体,诱导表达后亲和色谱纯化,对纯化表达产物进行一系列检测,考察其抑瘤作用。结果表达产物NDRG4蛋白相对分子质量为38 856,表达量为19.58%,纯度为92%,等电点为6.2,流式细胞术证实该蛋白质对HHCC和7901肿瘤细胞均阻滞在G1期,细胞实验和裸鼠致瘤抑制实验证实该蛋白质对肿瘤细胞的生长和肿瘤的发生发育有较强抑制作用。结论构建了NDRG4的PQE30/M15表达载体并取得高表达,表达蛋白质有较强抑瘤活性,为基因功能研究奠定了基础。
Purpose To clone and to express tumor inhibition gene NDRG4, to purify the expressed product and to study its tumor inhibitory effect.Methods NDRG4 gene was cloned into pQE30/M15 expression vector and expressed under induction of IPTG. Expressed protein was purified by affinity chromatography. The inhibitory effects of NDRG4 protein were studied with a series of examinations. Results The expressed products, with a relative molecular weight of 38 856 and an isoelectric point of 6.2, were up to 19.58 % of total cellular protein, and reached a purity of 92 % after purification. The expressed NDRG4 protein showed strong inhibitory effects on the tumor cells and the tissues. Conclusion An expression vector pQE30/M15 for NDRG4 was constructed, and NDRG4 protein showed strong tumor inhibition activity. It has laid a foundation for further study on genetic function of the protein.
出处
《中国生化药物杂志》
CAS
CSCD
2006年第3期137-141,共5页
Chinese Journal of Biochemical Pharmaceutics
关键词
NDRG4
克隆
表达
纯化
肿瘤抑制作用
NDRG4
cloning
expression
purification
tumor inhibition effect
