| 许婉芳,李祖剑.人TCTP基因的克隆与原核表达[J].井冈山大学自然版,2011,(6):46-50 |
| 人TCTP基因的克隆与原核表达 |
| CLONING AND EXPRESSION OF THE TRANSLATIONALLY CONTROLLED TUMOR PROTEIN(TCTP) IN HOMO SAPIENS |
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| DOI： |
| 中文关键词: 翻译控制肿瘤蛋白 重组 PCR 表达 纯化 |
| 英文关键词: translationally controlled tumor protein(TCTP) recombination,PCR expression purification |
| 基金项目:福建省教育厅A类科技项目(JA10236);泉州师范学院校级自选项目(2010KJ05) |
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| 中文摘要: |
| 人翻译控制肿瘤蛋白（hTCTP）是一个在进化过程中高度保守且具有很高表达量的蛋白家族。本实验通过PCR技术扩增人hTCTP基因,经酶切后插入表达质粒pGEX-4T-2构建表达载体,转化大肠杆菌后挑选阳性菌进行菌落PCR及质粒酶切鉴定,确证克隆成功并构建了人hTCTP基因表达载体。重组表达载体经IPTG诱导表达出目的蛋白,用超声破碎、亲和层析和SDS-PAGE对目的蛋白进行了分离纯化并鉴定。 |
| 英文摘要: |
| Human translationally controlled tumor protein(hTCTP) is a highly conserved protein family with high protein expression level during the evolution.The hTCTP gene was amplified using PCR in the experiment.Then the gene was inserted into the expression plasmid of pGEX-4T-2 after it was digested to build the expression vector.Colony PCR and enzyme digestion were used to confirm the successful cloning and construction of hTCTP expression vector.The recombination vector pGEX-4T-2-hTCTP was induced by IPTG,and the expression protein was separated,purified and identified using centrifugation,sonication,affinity chromatography and SDS-PAGE. |
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