HYPERACTIVE MUTANTS OF HIMAR1 TRANSPOSASE AND METHODS FOR USING THE SAME

摘要

Mariner-family transposable elements are active in a wide variety of organis ms and are becoming increasingly important genetic tools in species lacking sophisticat ed genetics. The Himar1 element, a member of the mariner family, isolated from the horn fly, Haematobia irritans, is active in Escherichia coli when expressed appropriately. Using this fact, a genetic screen was devised to isolate hyperactive mutants of Himar1 transposase that enhance overall transposition from 4 to 50-fold as measured in an E. coli assay. These hyperactive Himar1 mutant transposases should enable sophisticated analysis of the biochemistry of mariner transposition and should improve efficiency of a variety of genetic manipulations involving transposition in vivo and in vitro such as random mutagenesis or transgenesi s in a wide range of host cells than the transposable elements previously available.