融合蛋白sCAR-TSP-1原核表达纯化及诱导白血病细胞K562凋亡的研究
Prokaryotic Expression and Purification of sCAR-TSP-1, and Induced Apoptosis in leukemic cells k562
根据thrombospondin-1（TSP-1）4N1端氨基酸序列(RFYVVMWK)，以已有的腺病毒受体sCAR为模板，将8个氨基酸序列通过PCR扩增于sCAR之后，得到目的序列sCAR-TSP-1，连接到表达载体pQE30上，转化大肠杆菌M15后获得工程茵。该菌株经IPTG诱导后，高效表达出带有组氨酸标签以包涵体形式存在的融合蛋白sCAR-TSP-1。包涵体经过尿素变性溶解、PBS稀释复性、Ni离子亲和层析柱纯化，获得目的蛋白。SDS-PAGE分析表明，在26kD附近有一条明显的特异性蛋白条带。同时细胞实验结果表明融合蛋白sCAR-TSP-1对白血病细胞K562有明显凋亡作用。
according to the amino acid sequence of thrombospondin-1(RFYVVMWK), To the adenovirus receptor sCAR as template, 8 amino acid sequence amplified by PCR from sCAR，obtain the sequences of sCAR-TSP-1, connected to the expression vector pQE30, was transformed into E. coli M15,obtained the engineering bacteria. The strain induced by IPTG, efficient expression with histidine label in the form of inclusion body of the fusion protein sCAR-TSP-1.The inclusion body is dissolved by urea denaturation, PBS dilution refolding, Ni ion affinity chromatography purification, and obtain the target protein. SDS-PAGE analysis shows that, in the vicinity of 26kD has an apparent specific protein bands. At the same time, the experimental results show that the fusion protein sCAR-TSP-1in leukemia cell K562 has obvious apoptosis.
梁天祥、武虎、谌贺宽子、陈磊、唐斌
生物科学研究方法、生物科学研究技术生物工程学分子生物学
SP-1sCAR-TSP-1融合蛋白原核表达白血病细胞
SP-1fusion protein sCAR-TSP-1prokaryotic expressionleukemic cell
梁天祥,武虎,谌贺宽子,陈磊,唐斌.融合蛋白sCAR-TSP-1原核表达纯化及诱导白血病细胞K562凋亡的研究[EB/OL].(2011-11-22)[2025-08-11].http://www.paper.edu.cn/releasepaper/content/201111-357.点此复制
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