Identification And Analysis Of The Transposition Activity Of Mariner-like Elements From Phyllostachys Pubescens

Transposable element, also known as the skip factor, and its essence is a certain length of DNA sequences that can change their sites. Mariner-like elements (short title MLE) is a kind of DNA transposons widespread in animal and plant genomes.Our group has separated a full-length mariner-like transposase from Phyllostachys pubescens.1) A yeast assay was devised to determine whether the mariner-like, elements encoded an active transposase:the transposase CDS are linked to one yeast vector pAG413-gal, and the nonautonomous transposon sequences are linked to the other yeast expression vector pWL89a. Transformants containing both plasmids were selected on plates of selective medium. Colonies were picked after induced screening and active ones are tested by PCR amplification. We found that the mariner-like, transposon of Bambusoideae Subfamily cound transpose in yeast. Sequencing of each active strain, and compared with this locus in the original plasmid, revealed that excision of transposase was precise where without transposon footprints.2) Arabidopsis thaliana assay is adopted as the plant transgenic expression system, identified the expression of transposon activity in plants:both full-length mariner-like, transposase and nonautonomous transposon sequences are linked to the expression vector PCAMBIA1304-. Transformants were identified and the expression of the transposon in Arabidopsis thaliana was detected.To the basis of the separation of a full-length mariner-like transposase from Phyllostachys pubescens, we demonstrated its transposition activity in the yeast assay, which will lay a foundation for further construction of the highly active mariner-like elements and the development of the molecular tagging; At the same time, the study of transgene expression in Arabidopsis thaliana lay a basis for the future research about unique escision and genetic stability in Arabidopsis thallana genome, and it will provide a new genetic tool for building a large-scale plant mutant library and studying gene function, which will make it easy to isolate and identify more genome from Phyllostachys pubescens and accelerate the process of its genomics research.