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Sitedirected mutagenesis of tryptofan residues in the structure of the catalytic module of tyrosiltRNA synthetase Bos taurus

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


SUMMARY. To study the structural-dynamic and functional properties of the N-terminal catalytic module of Bos taurus tyrosyl-tRNA synthetase (mini BtTyrRS) by fluorescence spectroscopy site-directed mutagenesis of the protein with the replacement of three Trp residues with Ala residues in its structure was performed using the modified QuikChang method. In the process of sequential PCR reactions using the developed primers point substitutions of tryptophan codons TGG with alanine codons GCG were made within the cDNA sequence of the tyrosyl-tRNA synthetase catalytic module cloned in the expression plasmid pET-30a. As a result, mini BtTyrRS cDNAs were obtained within the nucleotide sequence of which there is only one codon of tryptophan in each of the three positions in the protein structure.

Key words: catalytic module of tyrosyl-tRNA synthetase, site-directed mutagenesis, cDNA, PCR amplification, DNA polymerase

Tsitologiya i Genetika 2019, vol. 53, no. 3, pp. 47-57

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E-mail: v.n.zayets, a.tsuvariev, babenko_lesia, kornelyuk

Zayets V.N., Tsuvarev A.Yu., Kolomiiets L.A., Kornelyuk A.I. Sitedirected mutagenesis of tryptofan residues in the structure of the catalytic module of tyrosiltRNA synthetase Bos taurus, Tsitol Genet., 2019, vol. 53, no. 3, pp. 47-57.

In "Cytology and Genetics":
V. N. Zayets, A. Yu. Tsuvarev, L. A. Kolomiiets, A. I. Kornelyuk Site-Directed Mutagenesis of Tryptophan Residues in the Structure of the Catalytic Module of Tyrosyl-tRNA Synthetase from Bos taurus, Cytol Genet., 2019, vol. 53, no. 3, pp. 219226
DOI: 10.3103/S009545271903006X


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