| 摘要 |
FIELD: molecular biology, genetic engineering, biochemistry, enzymes. SUBSTANCE: invention relates to preparing novel proteins and can be used in any analysis based on determination of adenosine triphosphate (ATP). Invention proposes mutant forms of enzyme luciferase with enhanced affinity to ATP preparing by replacing amino acid glutamate relating to conservative region sequence of enzyme of wild type and corresponding to residue in position 270 of luciferase from Photinus pyralis for other amino acid residues, in part, lysine, arginine, glutamine or alanine and addition of amino acid residue provides reduction of KM value. As result of addition of additional mutations involving the replace of glutamate residue at position corresponding to residue 354 in luciferase structure from Photinus pyralis for lysine residue and/or the replace of residue at position corresponding to residue 215 in structure of luciferase from Photinus pyralis for residue of hydrophobic amino acid the proposed mutant forms of enzyme confer the enhanced thermal stability additionally. Invention relates also to preparing DNA sequences encoding indicated mutant forms of luciferase and plasmid vector structures providing expression of corresponding recombinant proteins in transformed cells of E. coli. EFFECT: improved preparing method, valuable properties of luciferase and DNA. 14 cl, 6 dwg, 2 tbl, 2 ex |
