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18S rRNA基因siRNA表达质粒的构建和鉴定

Construction and identification of siRNA expression plasmid for interference buffalo 18S rRNA
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摘要 目的:构建和鉴定水牛18SrRNA干扰真核表达载体,为进一步研究18SrRNA功能奠定基础。方法:人工设计合成水牛18SrRNA-sh565干扰序列,经基因重组定向克隆插入到真核表达载体pGPH1/GFP/Neo,随机挑选2个克隆菌提取质粒并进行酶切和测序鉴定。结果:质粒酶切和测序结果显示,siRNA寡核苷酸按正确方向和序列插入质粒。结论:成功构建pGPH1/GFP/Neo-buffalo-18S rRNA-sh565干扰表达质粒。 Objective: To construct and identify siRNA expression plasmid for interference buffalo 18S rRNA. Methods: Designed buffalo-18S rRNA-sh565 oligonucletides were chemically synthesized and inserted into pGPH1/GFP/Neo vector after annealing, then transformed into E.Coli DH5α. The recombinant plasmid was identified by restriction endonuclease and DNA sequencing. Results: DNA sequencing showed the siRNA oligonucletides were correctly inserted into the eukaryotic expression vector pGPH1/GFP/Neo. Conclusion: siRNA expression plasmid for interference buffalo 18S rRNA is successfully constructed and lays the foundation for further study of 18S rRNA
出处 《海南医学院学报》 CAS 2006年第5期393-395,共3页 Journal of Hainan Medical University
基金 国家自然科学基金资助项目(NO:30360039)
关键词 RNA干扰 18S RRNA 基因表达 真核细胞 质粒 水牛 RNA interference 18S rRNA Gene expression Eukaryotic cell Plasmid Buffalo
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  • 1Saijou E, Fujiwara T, Suzaiki T, et al. RBD-1.a nueleolar RNA-binding protein, is essential for Caenorhabditis elegans early development through 18s ribosomal RNA processing[J], Nucleic Acids Res, 2004,32 ( 3 ) : 1028-1036.

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