TRANSLATION OF INSULIN-RELATED POLYPEPTIDES FROM MESSENGER-RNAS WITH TANDEMLY REITERATED COPIES OF THE RIBOSOME BINDING-SITE
TRANSLATION OF INSULIN-RELATED POLYPEPTIDES FROM MESSENGER-RNAS WITH TANDEMLY REITERATED COPIES OF THE RIBOSOME BINDING-SITE
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DOI：
10.1016/0092-8674(83)90554-8
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发表时间：
1983-01-01
期刊：
影响因子：
64.5
通讯作者：
KOZAK, M
中科院分区：
文献类型：
--
作者：
KOZAK, M
构建的质粒含有 66 bp [碱基对]片段的重复拷贝，大致称为核糖体结合位点，其中包括前胰岛素原的 AUG 起始密码子。极端测试涉及质粒 255/17，其携带核糖体结合位点的 4 个串联拷贝，所有 4 个 AUG 三联体与下游的前胰岛素原编码序列位于相同的阅读框中。在任何潜在起始位点的起始都会产生可被抗胰岛素抗血清沉淀的多肽，并且其大小将揭示起始中活性的AUG。 p255/17转染细胞中合成了一种胰岛素相关多肽；其大小对应于串联阵列中第一个核糖体结合位点起始的产物。由于未使用的 3 个下游 AUG 三联体出现在与所使用的 5''-近端 AUG 三联体周围相同的序列背景中，因此 AUG 三联体相对于 mRNA 5'' 端的位置对于将其识别为功能性起始密码子似乎很重要。
Plasmids were constructed containing reiterated copies of a 66 bp [base pair] fragment, loosely referred to as the ribosome binding site, that includes the AUG initiator codon of preproinsulin. The extreme test involved plasmid 255/17, which carried 4 tandem copies of the ribosome binding site, with all 4 AUG triplets in the same reading frame as the preproinsulin coding sequence downstream. Initiation at any potential start site would generate a polypeptide precipitable with anti-insulin antiserum, and its size would reveal the AUG(s) active in initiation. One insulin-related polypeptide was synthesized in cells transfected by p255/17; its size corresponded to the product initiated at the 1st ribosome binding site in the tandem array. Inasmuch as the 3 downstream AUG triplet, which are not used, occur in a sequence context identical with that around the 5''-proximal AUG triplets, which is used, the position of an AUG triplet relative to the 5'' end of the mRNA appears to be important in identifying it as a functional initiator codon.