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大肠杆菌酪氨酸抑制性转移核糖核酸前体结构基因的全合成。6. 对应于核苷酸序列100 - 126的脱氧核糖多核苷酸片段的合成。

Total synthesis of the structural gene for the precursor of a tyrosine suppressor transfer RNA from Escherichia coli. 6. Synthesis of the deoxyribopolynucleotide segments corresponding to the nucleotide sequence 100-126.

作者信息

Agarwal K L, Caruthers M H, Büchi H, van de Sande J H, Khorana H G

出版信息

J Biol Chem. 1976 Feb 10;251(3):624-33.

PMID:765331
Abstract

Chemical syntheses of the tridecanucleotide, d(G-C-T-T-C-C-C-G-A-T-A-A-G), the dodecanucleotide, d(G-C-T-C-C-C-T-T-A-T-C-G), the decanucleotide, d(G-G-A-G-C-A-G-G-C-C), the nonanucleotide, d(T-A-C-T-G-G-C-C-T), and the hexanucleotide, d(G-G-A-A-G-C), are described. Together, these syntheses represent the nucleotide sequence 100-126 of the DNA corresponding to the Escherichia coli tyrosine tRNA precursor. The synthesis of the dodecanucleotide d(G-C-T-C-C-C-T-T-A-T-C-G), was accomplished by the condensation of the previously described protected nonanucleotide, d[(MeOTr)ibG-anC-T-anC-anC-anC-T-T-bzA], with the trinucleotide block d[pT-anC-mbG(Ac)]. Synthesis of the other segments involved stepwise condensations to the 3'-OH group of growing oligonucleotide chains, starting with suitably protected deoxyribonucleosides and using protected mono-, di-, and trinucleotides as the incoming blocks. The final products, after deprotection, were purified by anion exchange chromatography and characterized. The synthesis of this part of the DNA was planned so that only a hexanucleotide segment is used to go up to the 3'-end (nucleotide 126) of the DNA and, therefore, it is amenable to elongation by chemical methods when the nucleotide sequence of the several nucleotides beyond this end becomes known.

摘要

描述了十三聚体核苷酸d(G-C-T-T-C-C-C-G-A-T-A-A-G)、十二聚体核苷酸d(G-C-T-C-C-C-T-T-A-T-C-G)、十聚体核苷酸d(G-G-A-G-C-A-G-G-C-C)、九聚体核苷酸d(T-A-C-T-G-G-C-C-T)和六聚体核苷酸d(G-G-A-A-G-C)的化学合成。这些合成共同代表了与大肠杆菌酪氨酸tRNA前体相对应的DNA的核苷酸序列100 - 126。十二聚体核苷酸d(G-C-T-C-C-C-T-T-A-T-C-G)的合成是通过将先前描述的受保护的九聚体核苷酸d[(MeOTr)ibG-anC-T-anC-anC-anC-T-T-bzA]与三核苷酸片段d[pT-anC-mbG(Ac)]缩合完成的。其他片段的合成涉及从适当保护的脱氧核糖核苷开始,以受保护的单核苷酸、二核苷酸和三核苷酸作为引入片段,逐步缩合到生长的寡核苷酸链的3'-OH基团上。脱保护后的最终产物通过阴离子交换色谱法纯化并进行表征。对该DNA这一部分的合成进行了规划,使得仅使用一个六聚体片段就可以延伸至DNA的3'-末端(核苷酸126),因此,当该末端以外的几个核苷酸的核苷酸序列已知时,它适合通过化学方法进行延伸。

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