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Cj1223c Mutant vs. Overexpressed, Time Course

ABSTRACT: The Cj1223c gene was cloned downstream of a strong promoter into the replicating plasmid pRY108, and was highly expressed in the wild type Campylobacter jejuni 81-176 strain (overexpressed). The Cj1223c gene was knocked out by allelic replacement in the 81-176 strain background (mutant). This mutant also carried an empty pRY108 vector. The Cj1223c overexpressing strain and the mutant were grown overnight in liquid broth, supplemented with kanamycin. The following morning, both cultures were diluted back to O.D.600=0.100, and allowed to continue to grow, shaking. At various time intervals (2, 6, 10, and 24 hours), samples of each culture were removed, combined with 1/10 stop solution, and the bacteria pelleted for RNA preparation. An all pairs experiment design type is where all labeled extracts are compared to every other labeled extract. Keywords: all_pairs

ORGANISM(S): Campylobacter jejuni

PROVIDER: GSE3198 | GEO | 2005/08/26

SECONDARY ACCESSION(S): PRJNA92861

REPOSITORIES: GEO

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Cj1223c Mutant vs. Overexpressed, Time Course

Project description:The Cj1223c gene was cloned downstream of a strong promoter into the replicating plasmid pRY108, and was highly expressed in the wild type Campylobacter jejuni 81-176 strain (overexpressed). The Cj1223c gene was knocked out by allelic replacement in the 81-176 strain background (mutant). This mutant also carried an empty pRY108 vector. The Cj1223c overexpressing strain and the mutant were grown overnight in liquid broth, supplemented with kanamycin. The following morning, both cultures were diluted back to O.D.600=0.100, and allowed to continue to grow, shaking. At various time intervals (2, 6, 10, and 24 hours), samples of each culture were removed, combined with 1/10 stop solution, and the bacteria pelleted for RNA preparation. An all pairs experiment design type is where all labeled extracts are compared to every other labeled extract. Computed

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