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由T4 RNA连接酶催化的具有特定序列的合成核糖三核苷酸的连接。

Joining of synthetic ribotrinucleotides with defined sequences catalyzed by T4 RNA ligase.

作者信息

Ohtsuka E, Nishikawa S, Fukumoto R, Tanaka S, Markham A F

出版信息

Eur J Biochem. 1977 Dec 1;81(2):285-91. doi: 10.1111/j.1432-1033.1977.tb11950.x.

DOI:10.1111/j.1432-1033.1977.tb11950.x
PMID:598370
Abstract

RNA ligase catalyzed the joining of pC-C-Ap with C-A-A in the synthesis of C-A-A-C-C-Ap, which has the sequence of the Escherichia coli tRNAfMet 3'-end. pC-C-A was also shown to be joined to C-A-A without any undesired self-polymerization. Joining of pC-C-A to various synthetic ribotriplets, such as C-C-A, A-A-A, C-C-C, U-U-U, U-A-G, C-C-G and U-U-C, was performed as well as joining to the partially substituted trimers with a photolabile o-nitrobenzyl group, C-Anbzl-A and C-C-Anbzl. The yields were C-A-A-C-C-A (69%), C-C-A-C-C-A (38%), A-A-A-C-C-A (66%), C-C-C-C-C-A (71%), U-U-U-C-C-A (50%), U-A-G-C-C-A (23%), C-C-G-C-C-A (43%) and U-U-C-C-C-A (46%). C-Anbzl-A was a slightly poorer acceptor than C-A-A and C-C-Anbzl did not serve as an acceptor. Recognition of acceptor molecules by RNA ligase is discussed in terms of affinity of oligonucleotides for the enzyme.

摘要

在合成具有大肠杆菌甲硫氨酸起始转运核糖核酸（tRNAfMet）3'端序列的C - A - A - C - C - Ap的过程中，RNA连接酶催化了pC - C - Ap与C - A - A的连接。还证明了pC - C - A能够与C - A - A连接，且没有任何不期望的自聚合现象。除了将pC - C - A与部分带有光不稳定邻硝基苄基的三聚物C - Anbzl - A和C - C - Anbzl连接外，还进行了pC - C - A与各种合成核糖三联体（如C - C - A、A - A - A、C - C - C、U - U - U、U - A - G、C - C - G和U - U - C）的连接。产率分别为C - A - A - C - C - A（69%）、C - C - A - C - C - A（38%）、A - A - A - C - C - A（66%）、C - C - C - C - C - A（71%）、U - U - U - C - C - A（50%）、U - A - G - C - C - A（23%）、C - C - G - C - C - A（43%）和U - U - C - C - C - A（46%）。C - Anbzl - A作为受体的能力略逊于C - A - A，而C - C - Anbzl不能作为受体。本文根据寡核苷酸对该酶的亲和力，讨论了RNA连接酶对受体分子的识别。

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