THE USE OF 5'-PHOSPHO-2 DEOXYRIBOCYTIDYLYLRIBOADENOSINE AS A FACILE ROUTE TO CHEMICAL AMINOACYLATION OF TRANSFER-RNA
THE USE OF 5'-PHOSPHO-2 DEOXYRIBOCYTIDYLYLRIBOADENOSINE AS A FACILE ROUTE TO CHEMICAL AMINOACYLATION OF TRANSFER-RNA
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DOI：
10.1093/nar/17.23.9649
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发表时间：
1989-12-11
影响因子：
14.9
通讯作者：
SCHULTZ, PG
中科院分区：
文献类型：
--
作者：
ROBERTSON, SA;NOREN, CJ;SCHULTZ, PG
描述了杂合二核苷酸5“-磷酸-2”-脱氧核糖胞苷酰核糖腺苷（pdCpA）的合成和化学氨酰化方法。使用T4 RNA连接酶将氨酰化pdCpA连接至截短的琥珀抑制基因tRNACUA（-CA）产生氨酰化抑制基因tRNA，其可用于将非天然氨基酸位点特异性掺入蛋白质中。当使用pCpA或pdCpA时，连接和体外抑制效率是相同的。脱氧胞苷的使用简化了合成二核苷酸pCpA的化学过程。此外，这些结果表明，在蛋白质合成过程中，核糖胞苷不需要识别氨酰化的tRNA。
Methodology is described for the synthesis and chemical aminoacylation of the hybrid dinucleotide 5''-phospho-2''deoxyribocytidylyriboadenosine (pdCpA). Ligation of aminoacylated pdCpA to a truncated amber suppressor tRNACUA (-CA) using T4 RNA ligase generates an aminoacylated suppressor tRNA which can be used for the site-specific incorporation of unnatural amino amino acids into proteins. Both the ligation and in vitro suppression effeciencies are the same when either pCpA or pdCpA is used. The use of deoxycytidine simplifies the chemistry involved in the synthesis of the dinucleotide pCpA. In addition, theses results demonstrate that ribocytidine is not required for recognition of the aminoacylated tRNA during protein synthesis.