| 引用本文： |
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马倩倩,童晓文.pIRES2-AcGFP1-sTRAIL真核表达载体的构建及鉴定[J].同济大学学报（医学版）,2011,32(2):35-38. [点击复制]
- MA Qian-qian,TONG Xiao-wen.Construction and identification of plRES2-AcGFPI-sTRAIL prokaryotic expression vector[J].同济大学学报（医学版）,2011,32(2):35-38. [点击复制]
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| 摘要: |
| 目的 利用基因工程技术合成sTRAIL（水溶性TRAIL）基因,并以AcGFP为报告基因,构建针对sTRAIL基因的pIRES2-AcGFP1-sTRAIL重组质粒。方法 从人胎盘组织中提取总RNA,利用RT-PCR方法扩增sTRAIL,并加入XhoI和BamHI酶切位点,通过双酶切将其连接于表达载体pIRES2-AcGFP1,通过PCR、酶切及测序鉴定重组质粒构建的正确性,最后用pIRES2-AcGFP1-sTRAIL重组质粒转染Hela细胞利用倒置荧光显微镜直接观察表达情况。结果 酶切、PCR及测序结果证实pIRES2-AcGFP1-sTRAIL构建序列正确。结论 利用基因工程技术成功构建真核表达载体pIRES2-AcGFP1-sTRAIL,为后续抗肿瘤研究奠定了基础。 |
| 关键词: TRAIL基因 真核表达载体 载体构建 |
| DOI：10.3969/j.issnl008-0392.2011.02.008 |
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| 基金项目: |
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| Construction and identification of plRES2-AcGFPI-sTRAIL prokaryotic expression vector |
| MA Qian-qian,TONG Xiao-wen |
| (Dept.of Gynecology and Obstetrics,Tongji Hospital,Tongji University,Shanghai 200065,China) |
| Abstract: |
| Objective To construct pIRES2-AcGFP1-sTRAIL prokaryotic expression vector.Methods The total RNA was extracted from the tissue of human placenta.The target fragment was amplified by RT-PCR,and then inserted into expression vector pIRES2-AcGFP1.The construct was identified by enzyme digestion,PCR,and sequencing.Results Endonuclease cutting,PCR,and sequencing showed the successful construction of pIRES2-AcGFP1-sTRAIL plasmid.Conclusions The extracellular region of sTRAIL gene is obtained successfully,laying a basis for its potential application to gene therapy. |
| Key words: TRAIL gene eukaryotic expression vector vector construction |
