Site-Directed Mutagenesis of Tryptophan Residues in the Structure of the Catalytic Module of Tyrosyl-tRNA Synthetase from Bos taurus

Zayets V. N.; Tsuvarev A. Yu.; Kolomiiets L. A.; Kornelyuk A. I.

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Site-Directed Mutagenesis of Tryptophan Residues in the Structure of the Catalytic Module of Tyrosyl-tRNA Synthetase from Bos taurus

机译：来自博斯金牛座的酪氨酰-CrNA合成酶催化模块结构中色氨酸残留的网站定向诱变

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Site-directed mutagenesis of the N-terminal catalytic module of Bos taurus tyrosyl-tRNA synthetase (mini-BtTyrRS) with the substitution of three Trp residues by Ala residues in its structure using the modified QuikChange method was performed to study structural dynamic and functional properties of the protein by fluorescence spectroscopy. Point substitutions of tryptophan codons TGG with alanine codons GCG in the cDNA nucleotide sequence of the tyrosyl-tRNA synthetase catalytic module cloned in expression plasmid pET-30a were obtained during sequential PCR reactions using the developed primers. As a result, mini-BtTyrRS cDNAs, whose sequences contain only one tryptophan codon in each of the three positions in the protein structure, were obtained.

机译：使用改性的Quikchange方法在其结构中通过Ala残基取代三次TRP残基的博士酪蛋杆菌酪氨酸-TrNA合成酶（Mini-BTTYRRS）的靶向诱变的诱变突变诱变，以研究结构动态和功能性荧光光谱法的蛋白质。在使用发育引物的顺序PCR反应期间获得了在表达质粒PET-30a中克隆的酪氨酰-TRNA合成酶催化模块的cDNA核苷酸序列中的色氨酸密码子GCG的点取代。结果，获得了Mini-Bttyrrs CDNA，其序列仅在蛋白质结构中的三个位置中的每一个中仅包含一个色氨酸密码子。

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来源
《Cytology and genetics》 |2019年第3期|219-226|共8页
作者
Zayets V. N.; Tsuvarev A. Yu.; Kolomiiets L. A.; Kornelyuk A. I.;
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Natl Acad Sci Ukraine Inst Mol Biol & Genet Kiev Ukraine;

Natl Acad Sci Ukraine Inst Mol Biol & Genet Kiev Ukraine|Taras Shevchenko Natl Univ Kyiv Kiev Ukraine;

Natl Acad Sci Ukraine Inst Mol Biol & Genet Kiev Ukraine;

Natl Acad Sci Ukraine Inst Mol Biol & Genet Kiev Ukraine;

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正文语种 eng
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catalytic module of tyrosyl-tRNA synthetase; site-directed mutagenesis; PCR amplification; DNA polymerase;

机译：酪氨酸-TRNA合成酶的催化模块;定点诱变;PCR扩增;DNA聚合酶;

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1. Site-Directed Mutagenesis of Tryptophan Residues in the Structure of the Catalytic Module of Tyrosyl-tRNA Synthetase from Bos taurus [J] . Zayets V. N., Tsuvarev A. Yu., Kolomiiets L. A., Cytology and genetics . 2019,第3期

机译：牛金牛座酪氨酰tRNA合成酶催化模块结构中色氨酸残基的定点诱变
2. Site-directed mutagenesis of catalytic residues in N ~5- Carboxyaminoimidazole ribonucleotide synthetase [J] . Dewal M.B., Firestine S.M. Biochemistry . 2013,第37期

机译：N〜5-羧氨基咪唑核糖核苷酸合成酶中催化残基的定点诱变
3. A brief perspective of the determination of crystal structures of site-directed mutants of tyrosyl-tRNA synthetase. [J] . Katherine A Brown Protein engineering design & selection: PEDS . 2011,第1a2期

机译：酪氨酸-tRNA合成酶定点突变体晶体结构测定的简要透视。
4. Probing the structure of human tissue factor by site-directed mutagenesis of tryptophan residues and in vim incorporation of tryptophan analogs [C] . C. A. Hasselbacher, Elena Rusinova, Evan Waxman, Fluorescence Science and Technology . 2000

机译：试抗鱼类残留物和vim掺入对色氨酸残基的人体组织因子结构的探讨
5. Site-directed mutagenesis defines tryptophan as the principal nitrosation site in human serum albumin. [D] . Harohalli, Krishna. 2000

机译：定点诱变将色氨酸定义为人血清白蛋白中的主要亚硝化位点。
6. Site-Directed Mutagenesis of Catalytic Residues in N5-Carboxyaminoimidazole Ribonucleotide Synthetase [O] . Mahender B. Dewal, Steven M. Firestine -1

机译：N5-羧氨基咪唑核糖核苷酸合成酶中催化残基的定点诱变
7. Isolation and characterization of the mutant form of N-terminal catalytically module of Bos taurus tyrosyl-tRNA synthetase with the replacement of Trp 40 and Trp 283 by alanine [O] . V. N. Zayets, D. M. Lozhko, O. Yu. Tsuvarev, 2020

机译：用丙氨酸替代TRP 40和TRP 283的Trp 40和Trp 283的N-末端催化模块的突变形式的突变形式的突变形式的突变形式

Site-Directed Mutagenesis of Tryptophan Residues in the Structure of the Catalytic Module of Tyrosyl-tRNA Synthetase from Bos taurus

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