Generation of Mutant
Isolation of Mutated Genomic DNA
Insertional Mutagenesis was Used to Generate a Non-Agglutinating Mutant
by transforming mt+ arg- cells with the pArg7.8 vector, which contains
the argininosuccinate lyase gene and thus rescues the arginine requirement.
Since the introduced DNA integrates randomly into the nuclear genome, each
integration event potentially disrupts a gene. Stable transformants were
selected on arginine-free media and then screened for their ability to mate
with a wild-type mt+ parent. In this way, a non-agglutinating mutant was
identified. Genomic DNA flanking the site of insertion was isolated by plasmid
rescue.