| Himar1 is a eukaryotic class II DNA-mediated transposon. It is a member of the mariner family of transposons, which are widespread in metazoans and are active in many organisms. Transposition of this element is carried out by a cut-and-paste mechanism, which is mediated by a self-encoded transposase and does not require any host-specific factors.;The Himar1 transposase binds DNA at specific inverted terminal repeat sequences at the transposon termini, excises the element from the donor DNA, and integrates at a TA dinuclotide at the target site. The entire mechanism of the transposition is dependent on the recognition and binding of the transposase protein to the inverted terminal repeat sequences.;This research was performed to determine the DNA-binding domain of the Himar1 transposase. The wild-type full-length Himar1 transposase was truncated by in vitro mutagenesis and screened using mobility shift DNA-binding analysis to map the DNA-binding domain of the Himar1 transposase protein. Further specific mutations were made within the mapped DNA-binding domain to test whether it affects the DNA-binding capabilities of the Himar1 transposase.;This structure and function study was carried out in order to understand the process of transposition and to be able to create hyperactive transposable elements, which can be harnessed as a better and much more efficient genetic tool. |
