Table 2

Effect of the AAV Rep protein expression on gene targeting and random integration in yeast
Plasmid Gene targeting Random integration GT/RI
ITR-lys2URA3lys2-ITR, No REP 31.8 ± 9.8 0.73 ± 0.39 43.5
ITR-lys2URA3lys2-ITR, REP 20.1 ± 5.9 0.88 ± 0.49 22.8
ITR-pokURA3-ITR, No REP ND 0.12 ± 0.08 /
ITR-pokURA3-ITR, REP ND 1.02 ± 0.56* /

Yeast was transformed with PvuII-restricted pAAVLUL or pAAVpokURA as reported in Materials and Methods. In the case of experiments with pAAVLUL, gene targeting and random integration events are reported as number of URA3+lys2-colonies /103 transformants per μg of plasmid DNA and URA3+LYS2+ colonies /103 transformants per μg of plasmid DNA, respectively. Similarly, in the case of pAAVpokURA experiments, non-homologous integration events are reported as total URA3+colonies /103 transformants per μg DNA. The number of transformants per μg of plasmid DNA was determined by transformation with 1 μg of episomal plasmid DNA. Results are the mean of four independent experiments ± standard deviation. The number of total colonies is indicated in the parentheses. Statistics was performed using Student’s t-test. *p = 0.0078 vsITR-pokURA3-ITR, No REP. ND = not detectable.

Galli and Cervelli

Galli and Cervelli BMC Molecular Biology 2014 15:5   doi:10.1186/1471-2199-15-5

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