人PSF蛋白结合长非编码RNA的筛选及鉴定  

Screening and identification of human polypyrimidine tract-binding protein-associated splicing factor(PSF)-binding long non-coding RNAs

作　　者：兰洋[1] 王慧娟[1] 刘涛[1] 李灵[1,2] 

机构地区：[1]四川大学生命科学学院功能基因组实验室,成都610064 [2]海南医学院海南省热带病重点实验室,海口571101

出　　处：《四川大学学报（自然科学版）》2011年第4期910-916,共7页Journal of Sichuan University(Natural Science Edition)

基　　金：国家自然科学基金(31000579);海南省自然科学基金(310045)

摘　　要：通过RNA亲和层析(RNA affinity chromatograph),4种可能与人PSF(human polypyrimidinetract-binding protein-associated splicing factor,hPSF)蛋白结合的长非编码RNA(1ong non-coding RNA,lncRNA)片段在人黑色素瘤细胞yusac细胞核RNA文库中被筛选得到.它们分别定位于人内源性逆转座子L1PA16、MER11C、非编码基因MALAT-1以及一个未知基因(unknown gene).紫外交联分析(UV cross-linking assay)证明在体外条件下这4种lncRNA片段均能与PSF蛋白结合;RNA免疫沉淀(RNA immunoprecipitation,RNA-IP)进一步证实在体内条件下这4种lncRNA均能与hPSF发生相互作用.最后,半定量RT-PCR表明,未知lncRNA特异高表达于人黑色素瘤细胞,其余3种lncRNA在多种人类肿瘤细胞中的丰度均较正常细胞明显升高.Four hPSF （human polypyrimidine tract-binding protein-associated splicing factor）-binding lncRNA （long non-coding RNA） fragments were isolated by RNA affinity chromatograph from a nuclear RNA library of human yusac melanoma cell line. They map to retrotransposon L1PA16 and MERllC, non-coding gene MALAT-1 and an unknown gene, respectively. UV cross-linking assay confirmed that the four lncRNAs bind to hPSF in vitro. RNA-immunoprecipitation （RNA-IP） demonstrated that they also form lncRNA/hPSF protein complexes in vivo. A screen of 9 human tumor cell lines by semi-quan titative RT-PCR shown that the lines. The other three lncRNAs cells. unknown lncRNA was exclusively expressed in human melanoma cell were also overexpressed in human tumor cells in contrast to normal

关 键 词：人PSF蛋白 长非编码RNA RNA亲和层析 

分 类 号：Q522[生物学—生物化学]