| 增强型绿色荧光蛋白RNAi表达载体的构建和鉴定 |
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| 引用本文： | 干惠珠,张桂珍,卜丽莎,杨绍娟,高申,郑德明.增强型绿色荧光蛋白RNAi表达载体的构建和鉴定[J].中国实验诊断学,2011,15(4):619-621. |
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| 作者姓名： | 干惠珠 张桂珍 卜丽莎 杨绍娟 高申 郑德明 |
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| 作者单位： | 1. 吉林大学中日联谊医院,肿瘤血液科,吉林,长春130033
2. 吉林大学中日联谊医院,中心研究室,吉林,长春130033 |
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| 摘 要： | 目的构建靶向增强型绿色荧光蛋白基因（EGFP）的RNA干扰质粒载体。方法设计并合成针对EGFP编码区的模板寡核苷酸单链,克隆入pSilencerTM 3.1-H1 neo载体中,对重组质粒进行酶切鉴定和DNA序列测定。结果重组质粒经限制性内切酶酶切得到66 bp和4.3 kb条带;重组阳性克隆测序结果与实验设计的模板寡核苷酸序列相符。结论成功构建靶向EGFP的RNA干扰表达载体,为RNA干扰技术在实验室的进一步开展奠定基础。
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| 关 键 词： | RNAi 增强型绿色荧光蛋白 shRNA表达载体 |
Construction and identification of RNA interference expression vector targeting EGFP |
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| GAN Hui-zhu,ZHANG Gui-zhen,BU Li-sa,et al..Construction and identification of RNA interference expression vector targeting EGFP[J].Chinese Journal of Laboratory Diagnosis,2011,15(4):619-621. |
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| Authors: | GAN Hui-zhu ZHANG Gui-zhen BU Li-sa |
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| Institution: | GAN Hui-zhu1,ZHANG Gui-zhen2,BU Li-sa2,et al.(Central Research Department,China-Japan Union Hospital,Jilin University,Changchun 130033,China) |
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| Abstract: | Objective To construct RNAi（RNA interference） expression vector targeting EGFP.Methods The oligos targeting EGFP were desigened and cloned into the BamHⅠand Hind Ⅲ site of linearized pSilencerTM 3.1-H1 neo shRNA expression vector.The recombinant plasmid vectors were confirmed by enzyme digestion analysis and DNA sequencing.Results The fragment of 66bp and 4.3kb was shown after digestion by BamHⅠ和Hind Ⅲ and agarose gel electrophoresis.,which was proved to be identical to original construction.Conclusion RNAi expression vector targeting EGFP has been constructed successfully,which facilitates the wider use of RNA interference technique in laboratory. |
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| Keywords: | RNAi |
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