In vitro-synthesized adenovirus 2 messenger RNA precursors are accurately spliced by nuclear extracts.
In vitro-synthesized adenovirus 2 messenger RNA precursors are accurately spliced by nuclear extracts.
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体外合成的腺病毒 2 信使 RNA 前体通过核提取物进行精确剪接。
DOI：
10.1073/pnas.81.15.4707
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发表时间：
1984
影响因子：
11.1
通讯作者：
Goldenberg,CJ
中科院分区：
文献类型：
--
作者：
Goldenberg,CJ
将腺病毒2的早期2区基因融合到有效的噬菌体启动子（沙门氏菌噬菌体6）的质粒中，体外合成了前体mrna。这些rna被纯化，并在MOPC-315小鼠骨髓瘤细胞核提取物的存在下作为底物进行体外剪接。体外剪接在核苷酸水平上是准确的。反应发生得很快，没有任何可察觉的迟滞。在孵育期间，前mrna前体的浓度似乎是体外RNA剪接效率高（60%-80%）的重要因素。剪接组分的分离以及DNA模板的修饰，以研究体外剪接所需的核苷酸序列，现在可以用该系统完成。
Precursor mRNAs were synthesized in vitro from a plasmid in which the early region 2 gene of adenovirus 2 is fused to an efficient bacteriophage promoter (Salmonella phage 6). The RNAs were purified and used as substrates for in vitro splicing in the presence of nuclear extracts prepared from MOPC-315 mouse myeloma cells. The in vitro splicing was accurate at the nucleotide level. The reaction occurs rapidly and without any detectable lag. The concentration of the pre-mRNA precursor during incubation appears to be an important factor for high efficiency (60%-80%) of in vitro RNA splicing. Fractionation of the splicing components as well as modifications of the DNA template to study the nucleotide-sequence requirement for in vitro splicing can now be accomplished with this system.