CC-2969 petA (HC-delta) spr sr mt+
$30.00
From Bruce Kohorn, Duke University, January 1994
Phenotype: antibiotic resistant (spectinomycin, streptomycin)
This is a site-directed mutation transformed into the signal sequence of the petA gene. CC-125 was co-transformed with P-183 (carrying sr-u-2-60 spr-u-1-6-2 er-u-37) and a plasmid carrying a site-directed mutation in the signal sequence for the petA gene. Transformants were selected for spectinomycin and streptomycin resistance; the er-u-37 marker was not tested. The original transformant grew very slowly.
HC[delta] is a hydrophobic core deletion made by creating two EcoRI sites through sequential in vitro mutagenesis of nucleotides 39-43 and nucleotides 61-66, subcloning the 2.3 kb KpnI fragment containing this modified petA gene into a pUC119 plasmid lacking both HindIII and EcoRI sites, digesting with EcoRI to remove the hydrophobic core, and then religating.
Smith TA, Kohorn BD (1994) Mutations in a signal sequence for the thylakoid membrane identify multiple protein transport pathways and nuclear suppressors. J Cell Biol 126:365-374