| 表达miR-150shRNA细胞株的初步建立 |
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| 引用本文： | 代晨,邓文,张彦明.表达miR-150shRNA细胞株的初步建立[J].动物医学进展,2009,30(8):1-5. |
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| 作者姓名： | 代晨 邓文 张彦明 |
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| 作者单位： | 西北农林科技大学动物医学院,陕西杨陵,712100 |
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| 基金项目： | 国家自然科学基金项目(30771607);;家畜疫病病原生物学国家重点实验室开放基金项目(猪瘟病毒感染相关microRNA的克隆与鉴定) |
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| 摘 要： | 为了探讨融合microRNA重组干扰载体构建的方法,为今后mi R-150对猪瘟病毒(CSFV)潜在的调控进行研究,利用生物信息学技术筛选出mi R-150,设计并合成了表达mi R-150的两条shRNA序列的DNA单链,将其退火后与干扰载体pGenesil-1连接,构建了含有mi R-150shRNA和绿色荧光蛋白基因的重组干扰质粒pGene-mi R-150shRNA,提取并纯化质粒后,采用脂质体法将重组质粒转染PK-15细胞,用G418抗性筛选。经过酶切鉴定和测序鉴定,表明成功构建了重组干扰质粒pGene-mi R-150shRNA,并在转染24 h后即可检测到绿色荧光。本研究成功得到稳定表达mi R-150的细胞株,所建的方法可以应用于各种microRNA的构建。
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| 关 键 词： | 猪瘟病毒 miR-150 发夹RNA 转染 |
Establishment of Cell Line Expressing MiR-150 shRNA |
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| DAI Chen,DENG Wen,ZHANG Yan-ming.Establishment of Cell Line Expressing MiR-150 shRNA[J].Progress In Veterinary Medicine,2009,30(8):1-5. |
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| Authors: | DAI Chen DENG Wen ZHANG Yan-ming |
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| Institution: | College of Veterinary Medicine;Northwest A & F University;Yangling;Shaanxi;712100;China |
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| Abstract: | To explore and establish a method of vector construction,and to research on potential regulation of CSFV by miR-150,miR-150 was selected by bioinformatics,two pairs of single DNA chain encoding short hairpin RNA(shRNA)of miR-150 were designed and synthesized,and were ligated to the interference vector pGenesil-1.The recombinant plasmid pGene-miR-150shRNA was constructed,and transfected into the PK-15 cells by liposome after extraction and purification.The cell line expressing miR-150 was established by scre... |
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| Keywords: | CSFV miR-150 short hairpin RNA transfection |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
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