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Complementation Analysis: A. tumefaciens C58 Proline

Complementation Analysis: A. tumefaciens C58 Proline. M9. *48 hours incubation at 37C *all strains grew as expected (#3 shows minimal breakthrough on primary lay with E. coli gene-likely due to leaky promoter) *Atu3985 proC complements; Atu2209 proC does not

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Complementation Analysis: A. tumefaciens C58 Proline

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  1. Complementation Analysis: A. tumefaciensC58 Proline M9 • *48 hours incubation at 37C • *all strains grew as expected • (#3 shows minimal breakthrough on primary lay with E. coli gene-likely due to leaky promoter) • *Atu3985 proC complements; Atu2209 proC does not • (the Atu2209 has been shown to complement, but much weaker than Atu3985) • *all strains tested on LB Km25; none grew • *all strains streaked 2x to purity prior to testing • (Exception for pkt-1, which was streaked 1x) WT 8005 #1 #5 M9 + 200 mg/ml proline M9 + 0.2% arabinose #3 #4c #4b #4a 8005 WT WT 8005 #1 #1 #5 #5 #3 #3 #4c #4c pKt-1= base vector + sacB 1522 bp pKt-3= base vector + E. coli proC 835 bp pKt-4= base vector + A. tumefaciens2209 proC 819 bp pKt-5= base vector + A. tumefaciens3985 proC 783 #4b #4a #4b #4a WT=E. coli K12 wild type (BW25113) 8005 = CGSC8005 proline auxotrophic E. coli K12 strain #1 = pKt-1 in 8005 (base vector + sacB) #3= pKt-3 in 8005 (E.coliK12 proC in base vector) #4a, #4b & #4c = pkt-4 in 8005; 3 clones; (Atu2209 proC in base vector) #5 = pKt-5 in 8005 (Atu3985 proC in base vector)

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