RNA-protein-interaction(完整)醫(yī)學(xué)課件

1、Protein-RNA InteractionsFunctional roles of RNA binding proteinsmRNA localizationmRNA translationmRNA splicingmRNA degradationmRNA editingMethods to study protein-RNA interactionEMSA (Electrophoretic mobility shift assays ) 蛋白質(zhì)與RNA結(jié)合后使其分子量增加RNA pull-down 下拉RNA，可同時獲得其結(jié)合的蛋白質(zhì)RIP (RNA immunoprecipitatio

2、n) 免疫沉淀蛋白質(zhì)，可同時獲得其結(jié)合RNACLIP-seq (Cross-Linking and ImmunoPrecipitation) 免疫沉淀蛋白質(zhì)，測序獲得其結(jié)合位點(diǎn)序列EMSA AUF1 binds to FST 3UTR1, 表達(dá)純化蛋白2, 蛋白和RNA結(jié)合3, 電泳RNA pull-downAUF1 binds to FST 3UTR through 112-221nt region1, PCR獲得biotin標(biāo)記的RNA片段1 4411 221222 4411 441codingbiotin2, biotin標(biāo)記的RNA片段與AUF1結(jié)合3, Streptavidin珠子下

3、拉RNA片段4, Western Blot檢測下拉產(chǎn)物中是否含有AUF1RIPRIP indenfies AUF1 binding with FST P21 and CyclinD1CLIP-seqTo study the binding sites of a specific proteinIntegrative Regulatory Mapping Indicates that the RNA-Binding Protein HuR Couples Pre-mRNA Processing and mRNA StabilityThe influence of HuR upon target

4、transcripts begins in the nucleus, where HuR associates with the pre-mRNA introns and affects splicing and likely other nuclear processing events. After assisting with export of the mature mRNA out of the nucleus, HuRs impact continues in the cytoplasm, where it stabilizes mRNAs, may help with transient mRNA storage, and modulates the re