Transposon-mediated site-specific recombination: A defined in vitro system

Research output: Contribution to journalArticle

Abstract

Transposition of the insertion element γδ is thought to involve formation of intermediates in which the element is present at each junction between donor and target replicons. In vivo these cointegrate structures are rapidly converted to the end products of transposition by site-specific recombination at a defined sequence, res, that is present in each directly repeated γδ element. Resolvase, an element encoded protein of molecular weight 21,000 is required for cointegrate resolution. I have demonstrated site-specific recombination in vitro using purified resolvase and a cointegrate analog substrate. The required components of the system described here are resolvase, negatively supercoiled substrate DNA, buffer and Mg2+. Neither host-encoded products nor high energy cofactors appear to be required for resolution in vitro. Catenated, resolved molecules are the major products of the reaction. Elimination of Mg2+ from the reaction yields different product molecules. The in vitro system described here provides an opportunity for detailed study of the resolution reaction.

Original languageEnglish (US)
Pages (from-to)713-719
Number of pages7
JournalCell
Volume25
Issue number3
DOIs
StatePublished - 1981
Externally publishedYes

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Recombinases
Genetic Recombination
Superhelical DNA
Replicon
Molecules
Substrates
Buffers
Molecular Weight
Molecular weight
DNA
In Vitro Techniques
Proteins

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

Transposon-mediated site-specific recombination : A defined in vitro system. / Reed, Randall R.

In: Cell, Vol. 25, No. 3, 1981, p. 713-719.

Research output: Contribution to journalArticle

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