Dave Eide wrote:
>Buddies,
>>We would like to do some one-step gene disruptions using flanking regions
>smaller than the recommended minimum. Rothstein (Meth. Enzymol. vol. 194)
>suggests using flanking regions of 100 bp or more on each side of the
selectable
>marker and the method we want to try gives us only about 50 bp on each side.
>Does anyone have information regarding homologous recombination of fragments of
>this size? Any input will be greatly appreciated.
>>Dave Eide
>It is possible to get targeting with sequences that short. See Baudin et al.
Nucl. Acid Res. 21: 3329 1993. Also, Fred Sherman had several papers a few
years ago describing transformation with oligos that restored function to
mutant cytochrome c genes. The efficiency of targeting to the intended site is
lower than you get with larger homologous regions.
Preston Garrison
Biochem. Dept.
UTHSCSA
San Antonio, Tx 78284-7760
USA
garrisonp at uthscsa.edu
