| pIRES2-AcGFP1-sTRAIL真核表达载体的构建及鉴定 |
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| 引用本文： | 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. DOI: 10.3969/j.issn.1008-0392.2011.02.008 |
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| 作者姓名： | 马倩倩 童晓文 |
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| 作者单位： | 同济大学附属同济医院妇产科,上海,200065 |
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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。
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| 关 键 词： | TRAIL基因 真核表达载体 载体构建 |
Construction and identification of plRES2-AcGFPI-sTRAIL prokaryotic expression vector |
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| MA Qian-qian and TONG Xiao-wen. Construction and identification of plRES2-AcGFPI-sTRAIL prokaryotic expression vector[J]. Journal of Tongji University(Medical Science), 2011, 32(2): 35-38. DOI: 10.3969/j.issn.1008-0392.2011.02.008 |
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| Authors: | MA Qian-qian and TONG Xiao-wen |
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| Affiliation: | (Dept.of Gynecology and Obstetrics,Tongji Hospital,Tongji University,Shanghai 200065,China) |
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| 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.Conclusion The extracellular region of sTRAIL gene is obtained successfully,laying a basis for its potential application to gene therapy. |
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| Keywords: | TRAIL gene eukaryotic expression vector vector construction |
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