Reaction Routes Enabled By Reversibly Acting Tyr-Recombinases and A Unidirectional Ser-Integrase
Fig. 2 shows, in sections A-B, the mode integration/resolution and inversion (INT/RES and INV) depend on the orientation of recombinase target sites (RTS), among these pairs of attP and attB. Section C indicates, in a streamlined fashion, the way recombinase-mediated cassette exchange (RMCE) can be reached by synchronous double-reciprocal crossovers (rather than integration, followed by resolution). Of note is the way reversible Flp-integration/resolution is modulated by 48 bp (in place of 34 bp minimal) FRT versions: the extra 13 bp arm serves as a Flp "landing path" contributing to the formation of the synaptic complex, both in the context of Flp-INT and Flp-RMCE functions (see the respective equilibrium situations). While it is barely possible to prevent the (entropy-driven) reversion of integration in section A for Cre and hard to achieve for Flp, RMCE can be completed if the donor plasmid is provided at an excess due to the bimolecular character of both the forward- and the reverse reaction. Posing both FRT sites in an inverse manner will lead to an equilibrium of both orientations for the insert (green arrow). In contrast to Flp, the Ser integrase PhiC31 (bottom representations) leads to unidirectional integration, at least in the absence of an recombinase-directionality (RDF-)factor. Relative to Flp-RMCE, which requires two different ("heterospecific") FRT-spacer mutants, the reaction partner (attB) of the first reacting attP site is hit arbitrarily, such that there is no control over the direction the donor cassette enters the target (cf. the alternative products). Also different from Flp-RMCE, several distinct RMCE targets cannot be mounted in parallel, owing to the lack of heterospecific (non-crossinteracting) attP/attB combinations.
Read more about this topic: Site-specific Recombinase Technology
Famous quotes containing the words reaction, routes, enabled and/or acting:
“Sole and self-commanded works,
Fears not undermining days,
Grows by decays,
And, by the famous might that lurks
In reaction and recoil,
Makes flames to freeze, and ice to boil.”
—Ralph Waldo Emerson (1803–1882)
“The myth of independence from the mother is abandoned in mid- life as women learn new routes around the mother—both the mother without and the mother within. A mid-life daughter may reengage with a mother or put new controls on care and set limits to love. But whatever she does, her child’s history is never finished.”
—Terri Apter (20th century)
““The unities, sir,’ he said, “are a completeness—a kind of universal dovetailedness with regard to place and time—a sort of general oneness, if I may be allowed to use so strong an expression. I take those to be the dramatic unities, so far as I have been enabled to bestow attention upon them, and I have read much upon the subject, and thought much.””
—Charles Dickens (1812–1870)
“But when I saw that they were not acting consistently with the truth of the gospel, I said to Cephas before them all, “If you, though a Jew, live like a Gentile and not like a Jew, how can you compel the Gentiles to live like Jews?””
—Bible: New Testament, Galatians 2:14.