MODIFIED TN5-TRANSPOSASE, SET FOR TRANSPOSITION, METHOD FOR TRANSPOSITION, DNA FRAGMENT AND GENETIC CONSTRUCTION OF MOBILE DNA SEQUENCE

摘要

FIELD: generic engineering, molecular biology, biochemistry. SUBSTANCE: invention describes a set for in vitro transposition involving DNA donor that carries mobile generic element flanked with pair of sequence- repeats at external ends of bacterial transposon Tn5, DNA-target to which mobile genetic element can be transferred and modified Tn5-transposase. Invention represents the modified Tn5- -transposase that with less probability as compared with Tn5-transposase of "wild" type confers inactive multidimensional formula. The modified transposase has substitution at position 54, 372 of Tn5-transposase of "wild" type and optionally at position 56. For transposition in vitro DNA-donor molecule is combined with DNA-target molecule and with modified Tn5-transposase. Invention describes DMA fragment encoding the modified Tn5-transposase. Genetic construction of mobile DNA sequence involves by it 5'- and 3'-ends the following flanking sequences: 5'-CTGTCTCTTATACACATCT-3' or 5'-CTGTCTCTT- -ATACAGATCT-3' compatible with the modified Tn5-transposase. Invention allows increasing the transposition reaction rate and to refuse in using external highly energetic source. EFFECT: valuable biological properties of set. 17 cl, 8 dwg, 1 tbl, 1 ex