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dc.contributor.author苏金华zh_CN
dc.contributor.author李文珠zh_CN
dc.contributor.author王生育zh_CN
dc.contributor.author黎之静zh_CN
dc.contributor.author陈彩霞zh_CN
dc.contributor.author庄国洪zh_CN
dc.date.accessioned2011-04-26T08:24:09Z
dc.date.available2011-04-26T08:24:09Z
dc.date.issued2008-11zh_CN
dc.identifier.citation免疫学杂志 Immunological Journal,2008,24(6):647-650zh_CN
dc.identifier.issn1000-8861zh_CN
dc.identifier.urihttps://dspace.xmu.edu.cn/handle/2288/8511
dc.description.abstract目的构建适于原核表达的重组蛋白RGD-FasL表达载体,并进行重组蛋白的表达纯化及抗肿瘤活性分析。方法通过重叠PCR将RGD序列插入到FasL基因的N端,获得RGD-FasL基因,构建pGEX-5X-1/RGD-FasL表达载体。转化大肠杆菌BL21(DE3),IPTG诱导表达,GST柱纯化。采用体外黏附实验、MTT比色法、流式细胞法检测融合蛋白的功能。结果通过重叠PCR获得了编码正确氨基酸序列的目的基因。目的蛋白以分泌的形式表达,表达量占菌体总蛋白的30%以上。纯化后,蛋白纯度达95%以上。体外黏附实验表明所纯化的融合蛋白可与宫颈癌Hela细胞发生特异结合。MTT比色法与流式细胞技术均表明纯化的融合蛋白能特异性地诱导肿瘤细胞发生凋亡。结论重组蛋白RGD-FasL表达载体的成功构建、表达、纯化及活性分析,为进一步的功能研究奠定了基础。 【英文摘要】 Objective To construct,express,and purify RGD-FasL gene and analysis its anti-tumor activities. Methods The RGD-FasL gene was constructed by overlapping PCR through inserting the sequence of RGD at the N-terminate of FasL gene,and then was inserted into vector pGEX-5X-1 and expressed efficiently in E.coli BL21(DE3) under optimization condition. The expressed products were purified by GST resin column. The function of recombined protein was detected by adhesion in vitro analysis,MTT colorimetric,and flow cyt...zh_CN
dc.language.isozhzh_CN
dc.publisher第三军医大学;中国免疫学会zh_CN
dc.subjectRGD-FasLzh_CN
dc.subject肿瘤zh_CN
dc.subject靶向治疗zh_CN
dc.subject凋亡zh_CN
dc.subjectTumorzh_CN
dc.subjectTarget therapyzh_CN
dc.subjectApoptosiszh_CN
dc.titleRGD-FasL基因的构建、表达、纯化及其活性分析zh_CN
dc.title.alternativeConstruction,expression,and purification of RGD-FasL and analysis of its activitieszh_CN
dc.typeArticlezh_CN


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