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.