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全合成一个序列与大肠杆菌甲酰甲硫氨酸转运核糖核酸相同的核糖核酸分子。

Total synthesis of a RNA molecule with sequence identical to that of Escherichia coli formylmethionine tRNA.

作者信息

Ohtsuka E, Tanaka S, Tanaka T, Miyake T, Markham A F, Nakagawa E, Wakabayashi T, Taniyama Y, Nishikawa S, Fukumoto R, Uemura H, Doi T, Tokunaga T, Ikehara M

出版信息

Proc Natl Acad Sci U S A. 1981 Sep;78(9):5493-7. doi: 10.1073/pnas.78.9.5493.

DOI:10.1073/pnas.78.9.5493
PMID:7029537
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC348772/
Abstract

A RNA molecule has been synthesized that is identical in sequence to Escherichia coli tRNAfMet except that it lacks the base modifications present in the E. coli tRNA. This was achieved by enzymatic joining of chemically synthesized oligonucleotides with chain lengths of 3-10 which were synthesized by the phosphodiester or phosphotriester method. First, quarter molecules of tRNA were constructed by joining of chemically synthesized fragments with RNA ligase. The 5'-quarter molecule (bases 1-20) served as an acceptor in joining reactions with the 3',5'-bisphosphorylated donor molecule (bases 21-34). The 5'-half molecule thus obtained was treated with phosphatase and joined to the 3'-half molecule which was prepared by ligation of the other quarter molecules (bases 35-60, acceptor; bases 61-77, donor) followed by 5'-phosphorylation with polynucleotide kinase. The synthetic tRNA was characterized by oligonucleotide pattern and was partially active in aminoacylation with E. coli methionyl-tRNA synthetase.

摘要

已合成一种RNA分子，其序列与大肠杆菌甲硫氨酸起始tRNA（tRNAfMet）相同，只是缺少大肠杆菌tRNA中存在的碱基修饰。这是通过将化学合成的链长为3至10的寡核苷酸进行酶促连接实现的，这些寡核苷酸是通过磷酸二酯法或磷酸三酯法合成的。首先，通过用RNA连接酶连接化学合成的片段构建tRNA的四分之一分子。5'-四分之一分子（碱基1至20）在与3'，5'-双磷酸化供体分子（碱基21至34）的连接反应中用作受体。将由此获得的5'-半分子用磷酸酶处理，并与3'-半分子连接，3'-半分子是通过连接其他四分之一分子（碱基35至60，受体；碱基61至77，供体）然后用多核苷酸激酶进行5'-磷酸化制备的。合成的tRNA通过寡核苷酸图谱进行表征，并且在用大肠杆菌甲硫氨酰-tRNA合成酶进行氨酰化反应中具有部分活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/49b8/348772/e06ee34078ae/pnas00660-0259-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/49b8/348772/1e525443b228/pnas00660-0258-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/49b8/348772/385d5d1c7dc6/pnas00660-0258-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/49b8/348772/4567a0544a81/pnas00660-0259-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/49b8/348772/e06ee34078ae/pnas00660-0259-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/49b8/348772/1e525443b228/pnas00660-0258-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/49b8/348772/385d5d1c7dc6/pnas00660-0258-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/49b8/348772/4567a0544a81/pnas00660-0259-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/49b8/348772/e06ee34078ae/pnas00660-0259-b.jpg

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Total synthesis of a RNA molecule with sequence identical to that of Escherichia coli formylmethionine tRNA.全合成一个序列与大肠杆菌甲酰甲硫氨酸转运核糖核酸相同的核糖核酸分子。
Proc Natl Acad Sci U S A. 1981 Sep;78(9):5493-7. doi: 10.1073/pnas.78.9.5493.
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本文引用的文献

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STRUCTURE OF A RIBONUCLEIC ACID.核糖核酸的结构
Science. 1965 Mar 19;147(3664):1462-5. doi: 10.1126/science.147.3664.1462.
2
DNA sequence analysis: a general, simple and rapid method for sequencing large oligodeoxyribonucleotide fragments by mapping.DNA序列分析：一种通过图谱对大的寡脱氧核糖核苷酸片段进行测序的通用、简单且快速的方法。
Nucleic Acids Res. 1974 Mar;1(3):331-53. doi: 10.1093/nar/1.3.331.
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Comparison of substrate base sequences for RNA ligase reactions in the synthesis of a tetradecanucleotide corresponding to bases 21-34 of E. coli tRNAfMet 1.
通过用与无义密码子UAG和UAA互补的三聚体替换大肠杆菌甲硫氨酸起始转运RNA（tRNAMetf）的反密码子三联体进行修饰。
Nucleic Acids Res. 1983 Jun 25;11(12):3863-72. doi: 10.1093/nar/11.12.3863.
4
Base substitutions in the wobble position of the anticodon inhibit aminoacylation of E. coli tRNAfMet by E. coli Met-tRNA synthetase.反密码子摆动位置的碱基替换会抑制大肠杆菌甲硫氨酰 - tRNA合成酶对大肠杆菌起始tRNAfMet的氨酰化作用。
Nucleic Acids Res. 1983 Mar 11;11(5):1439-55. doi: 10.1093/nar/11.5.1439.
5
E. coli initiator tRNA analogs with different nucleotides in the discriminator base position.在鉴别碱基位置具有不同核苷酸的大肠杆菌起始tRNA类似物。
Nucleic Acids Res. 1982 Oct 25;10(20):6531-9. doi: 10.1093/nar/10.20.6531.
6
Enzymatic replacement in vitro of the first anticodon base of yeast tRNAAsp: application to the study of tRNA maturation in vivo, after microinjection into frog oocytes.体外酶促置换酵母天冬氨酸tRNA的第一个反密码子碱基：在显微注射到蛙卵母细胞后用于体内tRNA成熟研究
Nucleic Acids Res. 1982 Jun 25;10(12):3715-32. doi: 10.1093/nar/10.12.3715.
7
Replacement and insertion of nucleotides at the anticodon loop of E. coli tRNAMetf by ligation of chemically synthesized ribooligonucleotides.通过化学合成的核糖寡核苷酸连接来替换和插入大肠杆菌甲硫氨酸起始转运核糖核酸（tRNAMetf）反密码子环处的核苷酸。
Nucleic Acids Res. 1985 May 24;13(10):3685-97. doi: 10.1093/nar/13.10.3685.
8
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9
Biochemical and physical characterization of an unmodified yeast phenylalanine transfer RNA transcribed in vitro.体外转录的未修饰酵母苯丙氨酸转移RNA的生化与物理特性
Proc Natl Acad Sci U S A. 1988 Feb;85(4):1033-7. doi: 10.1073/pnas.85.4.1033.
10
Chemical synthesis of a biologically active natural tRNA with its minor bases.具有稀有碱基的生物活性天然转运RNA的化学合成。
Nucleic Acids Res. 1992 Oct 11;20(19):5159-66. doi: 10.1093/nar/20.19.5159.
用于合成对应于大肠杆菌甲硫氨酸起始tRNA第21 - 34位碱基的十四聚核苷酸的RNA连接酶反应的底物碱基序列比较1。
Nucleic Acids Res. 1980 Sep 11;8(17):3909-16. doi: 10.1093/nar/8.17.3909.
4
Studies on transfer ribonucleic acids and related compounds. XXXI. Synthesis of the 3',5'-bisphosphorylated hexanucleotide corresponding to bases (72--77) of tRNAfMet from E. coli and its base transition analog.转移核糖核酸及相关化合物的研究。XXXI. 大肠杆菌甲硫氨酸起始转运核糖核酸（tRNAfMet）中对应碱基（72 - 77）的3',5'-双磷酸化六核苷酸及其碱基转换类似物的合成。
Chem Pharm Bull (Tokyo). 1980 Jan;28(1):120-5. doi: 10.1248/cpb.28.120.
5
Studies on transfer ribonucleic acids and related compounds. XXXVIII. A rapid method for the synthesis of ribooligonucleotides by using 3',5'-unsubstituted nucleosides. Synthesis of a hexanucleotide containing anticodon triplet of E. coli tRNA fMet.转移核糖核酸及相关化合物的研究。三十八。一种利用3'，5'-未取代核苷合成核糖寡核苷酸的快速方法。大肠杆菌甲硫氨酸起始转运核糖核酸反密码子三联体六核苷酸的合成。
Chem Pharm Bull (Tokyo). 1981 Mar;29(3):759-65. doi: 10.1248/cpb.29.759.
6
Studies on transfer ribonucleic acids and related compounds. XXXIV. Stepwise diester or partial triester synthesis of penta- to octanucleotides corresponding to residues 41--46, 47--54, 61--65 and 66--71 of tRNAfMet of E. coli.
Chem Pharm Bull (Tokyo). 1980 Aug;28(8):2450-9. doi: 10.1248/cpb.28.2450.
7
Synthesis of 5' fragments of formylmethionine transfer ribonucleic acid and their reconstitution with a natural three-quarter molecule.
Eur J Biochem. 1980 Apr;105(3):481-7. doi: 10.1111/j.1432-1033.1980.tb04523.x.
8
Chromatography of 32P-labelled oligonucleotides on thin layers of DEAE-cellulose.32P 标记的寡核苷酸在二乙氨基乙基纤维素薄层上的色谱分析。
Eur J Biochem. 1969 Dec;11(2):395-9. doi: 10.1111/j.1432-1033.1969.tb00786.x.
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Recovery of transfer RNA functions by combining fragmented Escherichia coli formylmethionine transfer RNA.通过组合片段化的大肠杆菌甲酰甲硫氨酸转运RNA恢复转运RNA功能
Biochim Biophys Acta. 1969 Oct 22;190(2):285-303. doi: 10.1016/0005-2787(69)90080-x.
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Purification of methionine-, valine-, phenylalanine- and tyrosine-specific tRNA from Escherichia coli.从大肠杆菌中纯化甲硫氨酸、缬氨酸、苯丙氨酸和酪氨酸特异性转运核糖核酸
Biochim Biophys Acta. 1967 Jun 20;142(1):133-48. doi: 10.1016/0005-2787(67)90522-9.