| 人前脑啡肽原基因真核表达载体的构建 |
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| 引用本文： | 马艳丽,赵青赞,曹靖,臧卫东,陈雪梅.人前脑啡肽原基因真核表达载体的构建[J].中原医刊,2007,34(15):3-4. |
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| 作者姓名： | 马艳丽 赵青赞 曹靖 臧卫东 陈雪梅 |
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| 作者单位： | [1]郑州大学第一附属医院麻醉科,郑州450052 [2]郑州大学医学院解剖教研室,郑州450052 |
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| 摘 要： | 目的 构建人前脑啡肽原基因真核表达载体，为进一步研究内源性脑啡肽的镇痛活性及相关药物研发奠定基础。方法 取新鲜的人脑颞叶皮质组织，用提取RNA的试剂盒提取总RNA，逆转录成cDNA。设计两对引物，用巢式PCR的方法获得人脑啡肽原基因，并将其插入到克隆载体pMD18-T中，克隆扩增。双酶切真核表达质粒pCDNA3．1（＋）及T载体中的目的片段，胶回收重组构建成pCDNA3．1（＋）-PENK表达载体。最后限制性酶切鉴定该表达载体。结果 采用巢式RT—PCR技术可以获得人前脑啡肽原基因，经测序证明其碱基序列为编码目的基因的正确序列。凝胶电泳结果证明已将此片段克隆到pCDNA3.1（＋）内。结论 成功构建了pCDNA3．1（＋）-PENK真核表达质粒。可作为疼痛基因治疗实验研究的有力工具。
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| 关 键 词： | 镇痛 脑啡肽 pCDNA3．1（＋）质粒 |
| 修稿时间： | 2007-04-06 |
Construction of recombinant pCDNA3.1(+)-PENK vector |
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| MA Yanli , ZHAO Qingzan, CAO Jing, ZANG Weidong, CHEN Xuemei.Construction of recombinant pCDNA3.1(+)-PENK vector[J].Central Plains Medical Journal,2007,34(15):3-4. |
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| Authors: | MA Yanli ZHAO Qingzan CAO Jing ZANG Weidong CHEN Xuemei |
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| Institution: | Department of Anesthesiology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan ; 2. Department of Anatomy,.Medical College of Zhengzhou University |
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| Abstract: | Objective To construct the recombinant pCDNA3.1(+)PENK vector encoding human enkephalin gene.Methods Human Enk gene was cloned by RT-PCR and inserted into pCDNA3.1(+)vector.Two pairs of primers were designed in which the restriction sites were compatible with the clone sites of the pCDNA3.1(+)vector.Using these primers Homosapiens proenkephalin(PENK)was amplified by polymerse chain reaction(PCR).Then it was inserted into the vector pcdna3.1(+).Results The recombinant retrovirus vector was constructed.Conclusion This recombinant plasmid may be a useful tool for gene therapy of pain and makes it possible to carry out further research in gene therapy of analgesic. |
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| Keywords: | Analgesic Enkephalin pCDNA3 1(+) |
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