Characterization of the newly-formed internucleotide bond of in vitro spliced mRNAs.
Characterization of the newly-formed internucleotide bond of in vitro spliced mRNAs.
复制标题
体外剪接 mRNA 新形成的核苷酸间键的表征。
DOI：
10.1016/0006-291x(84)90499-6
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发表时间：
1984
影响因子：
3.1
通讯作者：
Goldenberg,CJ
中科院分区：
文献类型：
--
作者：
Szeberènyi,J;Goldenberg,CJ
将腺病毒2的早期2区（E2）融合到有效的噬菌体启动子（沙门氏菌噬菌体6）的质粒中，体外合成前体rna。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。我们之前已经证明（Goldenberg， CJ， PNAS, August, in press, 1984），这些提取物的体外剪接在核苷酸水平上是准确的。我们现在证明：i)在体外剪接处产生的新的核苷酸间键是一个3 ',5 ' -磷酸二酯键；ii)形成外显子之间剪接的磷酸盐来源于pre-mRNA。
Precursor RNAs were synthesized in vitro from a plasmid in which the early region 2 (E2) of adenovirus 2 is fused to an efficient bacteriophage promoter (Salmonella phage 6). The RNAs were purified and utilized as substrates for in vitro splicing in the presence of nuclear extracts prepared from MOPC-315 mouse myeloma cells. We have shown previously (Goldenberg, CJ, PNAS, August, in press, 1984) that in vitro splicing in those extracts was accurate at the nucleotide level. We now show that: i) the new internucleotide bond at the splice junction generated in vitro is a 3′, 5′-phosphodiester bond; and ii) the phosphate that forms the splice between the exons is derived from the pre-mRNA.