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| miR-29c真核表达载体构建及鉴定 | |
| 引用本文： | 宋艳,张茂林,关振宏,段铭,张菁.miR-29c真核表达载体构建及鉴定[J].中国公共卫生,2012,28(9):1241-1243. |
| 作者姓名： | 宋艳 张茂林 关振宏 段铭 张菁 |
| 作者单位： | 1. 吉林大学人兽共患病研究所病毒室人兽共患病研究教育部重点实验室,吉林长春130062;北华大学护理学院 2. 吉林大学人兽共患病研究所病毒室人兽共患病研究教育部重点实验室,吉林长春,130062 3. 吉林大学人兽共患病研究所病毒室人兽共患病研究教育部重点实验室,吉林长春130062;湖南农业大学动物医学院 |
| 摘 要： | 目的构建pcDNA6.2-GW/EmGFP-miR-29c真核表达载体,为进一步研究miR-29c的功能和靶基因鉴定奠定基础。方法根据miR-29c成熟体序列设计合成1对miRNA寡聚单链DNA,经退火后克隆到真核表达载体pcDNA6.2-GW/EmGFP-miR中,瞬时转染肺腺癌A549细胞后,经实时荧光定量PCR检测pcDNA6.2-GW/EmGFP-miR-29c在肺癌细胞中表达miR-29c。结果 A549转染pcDNA6.2-GW/EmGFP-miR-29c 24 h后,miR-29c表达量上升,明显高于阴性对照,差异有统计学意义(P<0.05)。结论成功构建真核表达载体pcDNA6.2-GW/EmGFP-miR-29c,并能有效表达miR-29c。 |
| 关 键 词： | miR-29c 真核表达载体 基因表达 |
Construction and identification of eukaryotic expression vector of MiR-29c |
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| Institution: | SONG Yan*,ZHANG Mao-lin,GUAN Zhen-hong,et al(*Key Laboratory of Zoonoses of Ministry of Education,Institute of Zoonoses,Jilin University,Changchun,Jilin Province 130062,China) |
| Abstract: | Objective To construct eukaryotic vector of MiR-29c for the research of biological functions and target genes of miR-29c.Methods According to the mature sequence of miR-29c,a pair of oligodeoxynucleotides was designed,synthesized,annealed,and inserted into pcDNA6.2-GW/EmGFP vector.Then the vector was transfected into lung cancer A549 cells,and the expression of miR-29c was detected with real-time PCR.Results Twenty-four hours after the transfection of pcDNA 6.2-GW-EmGFP into A549 cells,the expression of miR-29c increased significantly compared to that of the negative control(P<0.05).Conclusion The recombinant pcDNA6.2-GW/EmGFP-miR-29c was successfully constructed. |
| Keywords: | miR-29c eukaryotic expression vector gene expression |
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