We present the crystal structure of the catalytic domain of Mos1 transposase, a member of the Tc1/mariner family of transposases. The structure comprises an RNase H-like core, bringing together an aspartic acid triad to form the active site, capped by N- and C-terminal alpha-helices. We have solved structures with either one Mg2+ or two Mn2+ ions in the active site, consistent with a two-metal ...

This communication reports the development of an efficient in vitro transposition system for Tn5. A key component of this system was the use of hyperactive mutant transposase. The inactivity of wild type transposase is likely to be related to the low frequency of in vivo transposition. The in vitro experiments demonstrate the following: the only required macromolecules for most of the steps in ...

The efficient production of transgenic (Tg) piglets has remained a challenge in the field domestic animal studies. Unlike mice, pronuclei pig zygotes cannot be easily studied because abundance lipid droplets. Therefore, must briefly centrifuged before pronuclear injection (PNI) to move droplets periphery zygote for PNI-mediated Tg piglets. However, this procedure is temporal return original spa...

Horizontal Gene Transfer (HGT) plays an important role in the physiology and evolution of microorganisms above all thermophilic prokaryotes. Some members of the Phylum Thermotogae (i.e., Thermotoga spp.) have been reported to present genomes constituted by a mosaic of genes from a variety of origins. This study presents a novel approach to search on the potential plasticity of Fervidobacterium ...

We used the Sleeping Beauty (SB) transposable element as a tool to probe transposon-host cell interactions in vertebrates. The Miz-1 transcription factor was identified as an interactor of the SB transposase in a yeast two-hybrid screen. Through its association with Miz-1, the SB transposase down-regulates cyclin D1 expression in human cells, as evidenced by differential gene expression analysi...

Insertion sequences (ISs) are ubiquitous and abundant mobile genetic elements in prokaryotic genomes. ISs often encode only one protein, the transposase, which catalyzes their transposition. Recent studies have shown that transposases of many different IS families interact with the β sliding clamp, a DNA replication factor of the host. However, it was unclear to what extent this interaction lim...

The Activator (Ac) element of maize is active at a low frequency in Arabidopsis. To determine whether this is due to poor expression of the Ac transposase gene, we obtained and studied 19 Arabidopsis transformants containing fusions of the octopine synthase (ocs), nopaline synthase (nos), cauliflower mosaic virus (CaMV) 35S, or Ac promoters to the transposase open reading frame. These transform...

The bacterial transposon Tn5 encodes two proteins, the transposase and a related protein, the transposition inhibitor, whose relative abundance determines, in part, the frequency of Tn5 transposition. The synthesis of these proteins is programmed by a complex set of genetic regulatory elements. The host DNA methylation function, dam, inhibits transposase promoter recognition and indirectly enha...

Synaptic complexes in prokaryotic transposons occur when transposase monomers bind to each of two specific end-binding sequences and then associate to bring the proteins and the two ends of the transposon together. It is within this complex of proteins and DNA that identical catalytic reactions are carried out by transposase on each of the ends of the transposon. In this study, we perform in vi...