| The focus of this thesis is the mechanistic study of non-LTR retrotransposon replication using the site specific transposon R2Bm. Non-LTR retrotransposons integrate into new chromosomal sites by copying their RNA back into DNA at the site of insertion using an exposed chromosomal 3' OH to prime cDNA synthesis.1 Although some of the early steps of an integration event have been detailed, how the element completes integration remains unknown.1 2 3 4 5 6 Clarifying the last integration step, known as second strand synthesis, is a major goal of this research. In order to accomplish this, R2 RNA and protein substrates were generated and used in biochemical assays to explore the aspects of RNP formation and second strand synthesis. |
