Abstract
Mariner-like elements (MLE) are a super-family of DNA transposons widespread in animal and plant genomes. Based on their transposition characteristics, such as random insertions and high-frequency heterogeneous transpositions, several MLEs have been developed to be used as tools in gene tagging and gene therapy. Two active MLEs, Ppmar1 and Ppmar2, have previously been identified in moso bamboo (Phyllostachys edulis). Both of these have a preferential insertion affinity to AT-rich region and their insertion sites are close to random in the host genome. In Ppmar2 element, we studied the affinities of terminal inverted repeats (TIRs) to DNA binding domain (DBD) and their influence on the transposition activity. We could identify two putative boxes in the TIRs which play a significant role in defining the TIR’s affinities to the DBD. Seven mutated TIRs were constructed, differing in affinities based on similarities with those of other plant MLEs. Gel mobility shift assays showed that the TIR mutants with mutation sites G669A-C671A had significantly higher affinities than the mutants with mutation sites C657T-A660T. The high-affinity TIRs indicated that their transposition frequency was 1.5–2.0 times higher than that of the wild type TIRs in yeast transposition assays. The MLE mutants with low-affinity TIRs had relatively lower transposition frequency from that of wild types. We conclude that TIR affinity to DBD significantly affects the transposition activity of Ppmar2. The mutant MLEs highly active TIRs constructed in this study can be used as a tool for bamboo genetic studies.
Highlights
Transposable elements (TEs) are genomic factors which can move around in the genome [1,2]
The terminal inverted repeats (TIRs) are usually 10–40 bp long and composed of protein binding elements (PBE) which are specific for individual mariner-like elements (MLE)
In the synthetic transposon Sleeping Beauty (SB), PBE is located in the TIRs of 15 bp long direct repeats (DR) [9]
Summary
Transposable elements (TEs) are genomic factors which can move around in the genome [1,2]. They have been employed as tools for genetic investigations in plants to improve their fitness and growth [3,4]. Among the Class II transposons, mariner-like elements (MLE) are a superfamily TEs widespread in diverse taxa, including higher animals, plants, fungi, insects, nematodes and fishes [6,7]. The TIRs are usually 10–40 bp long and composed of protein binding elements (PBE) which are specific for individual MLE. In the synthetic transposon Sleeping Beauty (SB), PBE is located in the TIRs of 15 bp long direct repeats (DR) [9]. FP and SB are Tc1/MLEs superfamily members sharing ~50% sequence similarity and used for gene therapy in fish, amphibian and mammalian cell lines [10,11]
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