Transposition of Tn5 in Escherichia coli is regulated by two transposon-encoded proteins: transposase (Tnp), promoting transposition preferentially in cis, and the trans-acting inhibitor (Inh). Two separate transposase mutants were isolated that replace glutamate with lysine at position 110 (EK110) and at position 345 (EK345). The EK transposase proteins increase the Tn5 transposition frequency...

studying vertebrate development, since the embryo could be a favorable target for gene transfer by in ovo electroporation in addition to classical manipulative advantages. However, it is still difficult to deliver transgenes into specific tissues at later developmental period, because common transfection methods are transient. We show here efficient transfer of EGFP driven by the GFAP promoter/...

studying vertebrate development, since the embryo could be a favorable target for gene transfer by in ovo electroporation in addition to classical manipulative advantages. However, it is still difficult to deliver transgenes into specific tissues at later developmental period, because common transfection methods are transient. We show here efficient transfer of EGFP driven by the GFAP promoter/...

studying vertebrate development, since the embryo could be a favorable target for gene transfer by in ovo electroporation in addition to classical manipulative advantages. However, it is still difficult to deliver transgenes into specific tissues at later developmental period, because common transfection methods are transient. We show here efficient transfer of EGFP driven by the GFAP promoter/...

The regulation of transcription and translation by specific cell types is essential to generate the cellular diversity that typifies complex multicellular organisms. Tagging and purification of ribosomal proteins has been shown to be an innovative and effective means of characterizing the ribosome bound transcriptome of highly specific cell populations in vivo. To test the feasibility of using ...

The transposon piggyBac is being used increasingly for genetic studies. Here, we describe modified versions of piggyBac transposase that have potentially wide-ranging applications, such as reversible transgenesis and modified targeting of insertions. piggyBac is distinguished by its ability to excise precisely, restoring the donor site to its pretransposon state. This characteristic makes piggy...

DNA transposons have been widely used for transgenesis and insertional mutagenesis in various organisms. Among the transposons active in mammalian cells, the moth-derived transposon piggyBac is most promising with its highly efficient transposition, large cargo capacity, and precise repair of the donor site. Here we report the generation of a hyperactive piggyBac transposase. The active transpo...

The effect of dam methylation on Tn5 transposition was investigated by analyses of mutations in the host (Escherichia coli) and the element. Wild-type elements transposed at a higher frequency and showed higher levels of transposase expression in a dam-host. Mutations were made in the promoter region of the transcript that codes for the transposase. Transposition and transposase levels from the...

Tn 5 is an excellent model system for understanding the molecular basis of DNA-mediated transposition. Mechanistic information has come from genetic and biochemical investigations of the transposase and its interactions with the recognition DNA sequences at the ends of the transposon. More recently, molecular structure analyses of catalytically active transposase; transposon DNA complexes have ...

Transgene insertion plays an important role in gene therapy and in biological studies. Transposon-based systems that integrate transgenes by transposase-catalyzed "cut-and-paste" mechanism have emerged as an attractive system for transgenesis. Hyperactive piggyBac transposon is particularly promising due to its ability to integrate large transgenes with high efficiency. However, prolonged expre...