使用5'-磷酸-2'-脱氧核糖胞苷酰核糖腺苷作为tRNA化学氨酰化的简便途径。
The use of 5'-phospho-2 deoxyribocytidylylriboadenosine as a facile route to chemical aminoacylation of tRNA.
作者信息
Robertson S A, Noren C J, Anthony-Cahill S J, Griffith M C, Schultz P G
机构信息
Department of Chemistry, University of California, Berkeley 94720.
出版信息
Nucleic Acids Res. 1989 Dec 11;17(23):9649-60. doi: 10.1093/nar/17.23.9649.
Abstract
Methodology is described for the synthesis and chemical aminoacylation of the hybrid dinucleotide 5'-phospho-2'-deoxyribocytidylylriboadenosine (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 site-specific incorporation of unnatural amino acids into proteins. Both the ligation and in vitro suppression efficiencies 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, these results demonstrate that ribocytidine is not required for recognition of the aminoacylated tRNA during protein synthesis.
摘要
本文描述了杂交二核苷酸5'-磷酸-2'-脱氧核糖胞苷酰核糖腺苷（pdCpA）的合成及化学氨酰化方法。使用T4 RNA连接酶将氨酰化的pdCpA连接到截短的琥珀色抑制tRNACUA（-CA）上，生成氨酰化抑制tRNA，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，连接效率和体外抑制效率相同。脱氧胞苷的使用简化了二核苷酸pCpA合成中涉及的化学过程。此外，这些结果表明，蛋白质合成过程中氨酰化tRNA的识别不需要核糖胞苷。
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