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.

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