Table 2.

Summary of PCR product transformation experiments

ExptRecipient strain (genotype)PCR product in transformationTotal no. of transformantsaNo. of integrants at wild-type allele of targeted locusScreening methodbComments
ARG5disruptionArg-het1 (arg5::hisG/ARG5)arg5::URA3182Argphenotype
ARG5 disruptionRM1000 (ARG5/ARG5)arg5::HIS1244Southern
Arg-het2 (arg5::HIS1/ARG5)arg5::URA3271Argphenotype & Southern21 transformants were His and Arg+
ADE2disruptionRM1000 (ADE2/ADE2)ade2::HIS1193Southern & PCR
Ade-het1 (ade2::HIS1/ADE2)ade2::URA3403Adephenotype & Southern12 transformants were His and Ade+
HRM101 disruptionBWP17 (HRM101/HRM101)hrm101::ARG4112PCR
Hrm-het1 (hrm101::ARG4/HRM101)hrm101::URA3142PCRArgtransformants were not screened
Hrm-het2 (hrm101::ARG4/HRM101)hrm101::URA3211PCRArgtransformants were not screened
ENX3disruptionBWP17 (ENX3/ENX3)enx3::URA352PCR
Enx-het1 (enx3::URA3/ENX3)enx3::ARG462PCRSelection eliminated Uri transformants
  • a Results from a single transformation are reported for each disruption.

  • b Details of screening methods are described in Materials and Methods and in Results.