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运用紫外交联免疫沉淀技术构建RNA结合蛋白的靶基因文库

Suwen Tian,Ruping Zhu

Genomics and Applied Biology（2017）

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Abstract
紫外交联免疫沉淀(crosslinking-immunoprecipitation,CLIP)是一种研究RNA结合蛋白的技术,它通过特异性抗体富集靶RNA片段,然后逆转录构建cDNA文库,最后进行高通量测序.它可以在基因组水平上全景式研究靶RNA的特点,为揭示该蛋白生物学功能与分子功能提供新的依据.本研究针对一个己知的RNA结合蛋白MOV 10 (moloney leukemia virus 10),通过CLIP技术构建cDNA文库,然后小规模克隆后测序,得到小部分靶RNA信息,并利用RNA免疫沉淀技术(RNA-immunoprecipitation,R1P)进行验证.测序提示MOV10结合mRNA,而其中有部分是其己知靶标.说明该文库可以用于后续深度测序,为研究该蛋白在精子发生领域内的功能和机制奠定基础.
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Key words
RNA binding protein,UV crosslinking-immunoprecipitation,Sequencing
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