3 Results
for term "sites"
- The 2009 Novitski Prize...of the plasmid integrating at the homologous site on a chromosome (Orr-Weaver et al. 1981, 1983). These experiments provided much of the impetus for developing a comprehensive double-strand-break-repair model of recombination (Szostak et al. 1983). But Rothsteins interest was also focused on methods to create ~~~
- The 2009 Thomas Hunt Morgan Medal...by Cairns et al. (1988). This publication suggested that selective stress might induce mutations or even direct them to sites that improve fitness. Roths lab in collaboration with the Swedish labs of Dan Andersson and Diarmaid Hughes has published substantial evidence that growth limitation enhances ~~~
Figure 1.—Ends-out gene targeting. (A) In budding yeast, a linearized DNA fragment containing a selectable marker in place of an open reading frame (ORF) recombines with homologous 5′ and 3′ sequences to create a full gene deletion. (B) In Drosophila, to create a targeted gene mutation requires several steps: (i) A modified P element (shaded box) is created containing FRT sites (solid arrows) surrounding 3′ and 5′ homologous sequences that flank all or part of the ORF. The homologous sequences are separated by an 18-bp I-SceI endonuclease recognition site (inverted triangles). (ii) Flies containing the P element are crossed with flies containing constructs that express both the site-specific FLP recombinase and the I-SceI endonuclease. FLP first pops out a circular DNA fragment and then I-SceI linearizes the excised segment. (iii) The linear fragment has an “ends-out” configuration that can replace part of the ORF with other sequences.