Tn552 transposase catalyzes concerted strand transfer in vitro
AUTOR(ES)
Leschziner, Andres E.
FONTE
The National Academy of Sciences
RESUMO
The Tn552 transposase, a member of the DDE superfamily of transposase and retroviral integrase proteins, has been expressed in soluble form. The purified protein performs concerted strand transfer in vitro, efficiently pairing two preprocessed transposon ends and inserting them into target DNA. For maximum efficiency, both participating DNA ends must contain the two adjacent transposase-binding sites that are the normal constituents of the Tn552 termini. As is the case with transposition in vivo, the insertions recovered from the reaction in vitro are flanked by repeats of a short target sequence, most frequently 6 bp. The reaction has stringent requirements for a divalent metal ion. Concerted strand transfer is most efficient with Mg2+. Although it stimulates strand transfer overall, Mn2+ promotes uncoupled, single-ended events at the expense of concerted insertions. The simplicity and efficiency of the Tn552 transposition system make it an attractive subject for structural and biochemical studies and a potentially useful genetic tool.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=22612Documentos Relacionados
- Tn552 transposase purification and in vitro activities.
- Frequency of Disinfectant Resistance Genes and Genetic Linkage with β-Lactamase Transposon Tn552 among Clinical Staphylococci
- In vitro transposition of Tn552: a tool for DNA sequencing and mutagenesis.
- p53 binds single-stranded DNA ends and catalyzes DNA renaturation and strand transfer.
- Identification of Genes Encoding Exported Mycobacterium tuberculosis Proteins Using a Tn552′phoA In Vitro Transposition System
