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GST-Glypican 3 N端融合蛋白表达载体构建及表达纯化 被引量：3

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摘要 目的构建人磷酯酰肌醇蛋白聚糖3(Glypican3,GPC3)N端蛋白与谷胱甘肽-S-转移酶(GST)重组的融合蛋白原核表达载体并进行表达及纯化。方法以重组质粒pcDNA3.1-GPC3N为模板,扩增GPC3 N端基因,产物经纯化回收后与载体PGEX-4T-1连接并转化大肠埃希菌Rosetta,酶切鉴定序列完全正确者诱导表达GST-GPC3 N端融合蛋白,并以谷胱甘肽琼脂糖小珠亲和纯化。结果Glypican 3 N端基因片段成功插入载体PGEX-4T-1,插入位点及碱基序列完全正确,转化Rosetta后,经IPGT诱导成功表达分子质量为64 000的GST-GPC3 N端融合蛋白。结论成功建立了重组GST-GPC3 N端融合蛋白的原核表达载体、表达菌株及诱导表达和纯化的方法,为GPC3 N端融合蛋白的进一步应用打下基础。
作者 肖明兵 赵敏星 倪润洲 江枫 周嘉伟
机构地区 南通大学附属医院消化病研究室 中国科学院上海神经科学研究所
出处 《临床检验杂志》 CAS CSCD 北大核心 2010年第3期212-214,共3页 Chinese Journal of Clinical Laboratory Science
基金 南通市社会发展计划项目(S7943)
关键词 磷酯酰肌醇蛋白聚糖3 载体 克隆 表达
分类号 Q78 [生物学—分子生物学]
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参考文献6

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共引文献2

同被引文献30

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引证文献3

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临床检验杂志
2010年 第3期

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