| 线粒体加工肽酶干扰RNA真核表达载体的构建 |
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| 引用本文： | 张艳梅,呼格吉乐,马飞煜,邬伟,马翔凌.线粒体加工肽酶干扰RNA真核表达载体的构建[J].中风与神经疾病杂志,2010,27(1). |
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| 作者姓名： | 张艳梅 呼格吉乐 马飞煜 邬伟 马翔凌 |
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| 作者单位： | 内蒙古医院神经内科,内蒙,呼和浩特,014000 |
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| 基金项目： | 内蒙古自治区自然基金资助项目(20080404MS1114) |
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| 摘 要： | 目的利用干扰RNA技术构建线粒体加工肽酶(mitochondrial processing peptidases,MPPs)干扰RNA真核表达载体,探讨MPPs在神经细胞中的作用,为帕金森病的研究提供有价值的资料。方法(1)根据Gen-Bank提供的大鼠MPPsα亚基(peptidase mitochondrial processing alpha,Pmpca)序列,设计并合成4对MPPs干扰RNA单链寡核苷酸:MPPs-SR-1113,MPPs-SR-1425,MPPs-SR-561,MPPs-SR-903。(2)使互补的单链寡核苷酸模板退火,再转化为能表达其小发卡结构RNA的DNA序列。(3)使PGPU6/GFP/Neo载体的线性化。(4)shRNA与载体的连接。(5)酶切鉴定并测序鉴定结果。结果(1)酶切结果表明,所有质粒均为阳性重组载体。(2)经鉴定证实,构建的siRNAs序列与基因库中序列完全相同,并且未发现有突变、缺失、插入等异常存在。结论利用干扰RNA技术可成功构建MPPs干扰RNA真核表达载体。
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| 关 键 词： | 干扰RNA 线粒体加工肽酶 帕金森病 载体 |
The construction of eukaryotic expression vector of RNA interference specific for mitochondrial processing peptidases |
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| ZHANG Yan-mei,HU Gejile,MA Fei-yu,et al..The construction of eukaryotic expression vector of RNA interference specific for mitochondrial processing peptidases[J].Journal of Apoplexy and Nervous Diseases,2010,27(1). |
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| Authors: | ZHANG Yan-mei HU Gejile MA Fei-yu |
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| Affiliation: | ZHANG Yan-mei,HU Gejile,MA Fei-yu,et al.(Neurology Department of Inner Monogolia Hospital,Huhhot 014000,China) |
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| Abstract: | Objective Eukaryotic expression vector of RNA interference specific for mitochondrial processing peptidases(MPPs)was constructed.It is critical to research the role that MPPs play in the nerve cells and provide valuable theoretical basis for the parkinsonism.Methods(1)Four pairs of siRNAs of MPPs gene were designed according to the principle of RNA interference:MPPs-SR-1113,MPPs-SR-1425,MPPs-SR-561,MPPs-SR-903.(2)Complementary single and strand DNA oligos was annealed and convertred into cDNA coding express... |
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| Keywords: | Interference RNA MPPs Parkinsonism Vector |
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