title: PLE-wu, a new member of piggyBac transposon family from insect, is active in mammalian cells.
abstract: piggyBac, a highly active transposon in insect and mammalian cells, is a very useful tool in genome manipulation. A new piggyBac-like element (PLE), named PLE-wu, was identified from a mutant baculovirus cultured in sf9 insect cells. This new transposon is 2931 bp in length and encodes two active forms of transposase, a 708-amino acid-long transposase and a short 576-residue-long transposase translated from a downstream in-frame initiation codon. PLE-wu has asymmetric terminal structures, containing 6-bp inverted terminal repeats, 32-bp imperfect inverted and direct sub-terminal repeats. Similar to piggyBac, PLE-wu exhibits traceless excision activity in both insect and mammalian cells, restoring the original TTAA target sequence upon excision. It also retains the insertion activity in mammalian cells with a plasmid to chromosome transposition rate about 10-fold higher than random integration. Plasmid rescue assays revealed that the TTAA target sequence was duplicated at the junctions of the insertion site. Deletion of the terminal sequences including the sub-terminal repeats decreased the transposition activity of the 708-residue-long transposase, while the transposition activity of the short form of transposase was not affected. With its low sequence similarity to piggyBac, PLE-wu will contribute to the understanding the mechanism of PLE transposition, as well as design of new transposon systems with higher activity.
authors.affiliation: Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, no. 04-01, Singapore 138669. Electronic address: cxwu@ibn.a-star.edu.sg.
authors.forename: Chunxiao
authors.initials: C
authors.lastname: Wu
authors.affiliation: Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, no. 04-01, Singapore 138669; Department of Biological Sciences, National University of Singapore, Singapore 117543.
authors.forename: Shu
authors.initials: S
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003062:Codon; D004251:DNA Transposable Elements; D016679:Genome, Viral; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010957:Plasmids; D017384:Sequence Deletion; D061987:Sf9 Cells; D018411:Spodoptera / Q000166:cytology / Q000821:virology; D019895:Transposases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D003062:Codon; D004251:DNA Transposable Elements; D014764:Viral Proteins; D019895:Transposases
keywords: Baculovirus; Insect; Mammalian; Transposon; piggyBac