所有细胞信使RNA的一个共同结构特征是poly(A) tail,它可以通过DNA模板(基于质粒或PCR)共转录引入,也可以通过转录后的酶促过程添加到mRNA中。含有poly(A)区域的质粒在大肠杆菌中重组导致poly(A)尾部广泛缩短。采用分段poly(A)方法我们可以显著减少大肠杆菌中质粒的重组而不会对mRNA半衰期和蛋白质表达产生负面影响。这...
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Understanding the function of poly(A) tails has been challenging because of difficulties in measuring their lengths in a genome-wide manner. Bartel and colleagues now report a new high-throughput sequencing method, called PAL-seq ('poly(A)-tail profiling by sequencing'), that accurately measures...
Poly(A)-tailprofilingrevealsan embryonicswitchintranslationalcontrol AlexanderO.Subtelny 1,2,3,4 *,StephenW.Eichhorn 1,2,3 *,GraceR.Chen 1,2,3 ,HazelSive 2,3 &DavidP.Bartel 1,2,3 Poly(A)tailsenhancethestabilityandtranslationofmosteukaryoticmessengerRNAs,butdifficultiesingloballymea- suringpoly...
1真核生物核基因转录生成的RNA前体经过修饰加工,在5'端加上5'cap;在3'端加上poly-Atail,再通过核孔进入细胞质中完成翻译过程,部分过程如图所示。基因A基因B基因C RNA前体13,5端修饰IRNA5'cap-poly- A tail加工5'cap poly- A tail(1)基因C对基因A的表达___(填“是”或“否”)有影响,理由是___。...
Poly(A) tails enhance the stability and translation of most eukaryotic mRNAs, but difficulties in globally measuring poly(A)-tail lengths have impeded greater understanding of poly(A)-tail function. Here, we describe poly(A)-tail length profiling by sequencing (PAL-seq) and apply it to measur...
近日,中国科学院遗传与发育生物学研究所陆发隆团队与东北林业大学刘玉胜团队、山东大学生殖医学研究中心陈子江/赵涵/吴克良团队、中国科学院动物研究所周兵团队以及东北农业大学王加强团队合作在Nature Structural & Molecular Biology发表了一篇题为Remodeling of maternal mRNA through poly(A) tail orchestrates human oocyte...
Purified recombinant AtRrp41p displays a processive phosphorolytic exonuclease activity and requires a single-stranded poly(A) tail on a substrate RNA as a loading pad. The expression of the Arabidopsis RRP41 cDNA in yeast rescues the 5.8 S rRNA processing and 3'-->5' mRNA degradation ...
常用的方法包括TAIL-Seq和PAL-Seq,借助于制备完整的包括polyA尾在内的3’端mRNA测序文库。首先将RNA与生物素化的3’端DNA接头连接,并用RNase T1部分酶切,留下完整的polyA尾,随后使用链霉亲和素磁珠捕获3’端片段。通过凝胶电泳筛选不同条带,并与5’端接头连接,为逆转录、文库扩增和测序提供完整的模板。如图3所...
本研究通过直接RNA测序(dRNA-seq)等一系列实验证明LARP1有助于在氨基酸饥饿(AAS)下具有长poly(A)尾部的TOP mRNA的选择性积累,从而加速核糖体加载到TOP mRNAs上,以便在AAS后恢复翻译。 英文标题:mTOR- and LARP1-dependent regulation of TOP mRNA poly(A) tail and ribosome loading ...