title: Transposition of the bamboo Mariner-like element Ppmar1 in yeast.
abstract: The moso bamboo genome contains the two structurally intact and thus potentially functional mariner-like elements Ppmar1 and Ppmar2. Both elements contain perfect terminal inverted repeats (TIRs) and a full-length intact transposase gene. Here we investigated whether Ppmar1 is functional in yeast (Saccharomyces cerevisiae). We have designed a two-component system consisting of a transposase expression cassette and a non-autonomous transposon on two separate plasmids. We demonstrate that the Ppmar1 transposase Pptpase1 catalyses excision of the non-autonomous Ppmar1NA element from the plasmid and reintegration at TA dinucleotide sequences in the yeast chromosomes. In addition, we generated 14 hyperactive Ppmar1 transposase variants by systematic single amino acid substitutions. The most active transposase variant, S171A, induces 10-fold more frequent Ppmar1NA excisions in yeast than the wild type transposase. The Ppmar1 transposon is a promising tool for insertion mutagenesis in moso bamboo and may be used in other plants as an alternative to the established transposon tagging systems.
authors.affiliation: The Nurturing Station for the State Key Laboratory of Subtropical Silviculture, Zhejiang A & F University, LinAn, China.
authors.forename: Ming-Bing
authors.initials: MB
authors.lastname: Zhou
authors.affiliation: The Nurturing Station for the State Key Laboratory of Subtropical Silviculture, Zhejiang A & F University, LinAn, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Hu
authors.affiliation: Paul Ehrlich Institute, Paul Ehrlich Str. 51-59, 63225 Langen, Germany.
authors.forename: Csaba
authors.initials: C
authors.lastname: Miskey
authors.affiliation: Institute of Biology, Dahlem Centre of Plant Sciences, Freie Universität Berlin, 14195 Berlin, Germany.
authors.forename: Katina
authors.initials: K
authors.lastname: Lazarow
authors.affiliation: Paul Ehrlich Institute, Paul Ehrlich Str. 51-59, 63225 Langen, Germany.
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Ivics
authors.affiliation: Institute of Biology, Dahlem Centre of Plant Sciences, Freie Universität Berlin, 14195 Berlin, Germany.
authors.forename: Reinhard
authors.initials: R
authors.lastname: Kunze
authors.affiliation: Department of Biology, University of Toronto, Mississauga, ON, Canada.
authors.forename: Guojun
authors.initials: G
authors.lastname: Yang
authors.affiliation: Max Delbrück Center for Molecular Medicine in the Helmholtz Society, Berlin, Germany. Electronic address: zizsvak@mdc-berlin.de.
authors.forename: Zsuzsanna
authors.initials: Z
authors.lastname: Izsvák
authors.affiliation: The Nurturing Station for the State Key Laboratory of Subtropical Silviculture, Zhejiang A & F University, LinAn, China. Electronic address: tangzafu@163.com.
authors.forename: Ding-Qin
authors.initials: DQ
authors.lastname: Tang
mesh_terms: D004251:DNA Transposable Elements; D018744:DNA, Plant; D010802:Phylogeny; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D031787:Sasa / Q000235:genetics*; D019895:Transposases / Q000378:metabolism
chemical_list: D004251:DNA Transposable Elements; D018744:DNA, Plant; D019895:Transposases
keywords: Mariner-like element; Moso bamboo (Phyllostachys edulis (Carrière) J. Houz.); Mutagenesis; Transposase