Sized-based marker identification technology

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving viable micro-organism

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435 6, 4351723, 4352351, 4352523, 43525411, 435325, 435419, C12Q 102, C12Q 168, C12N 1509, C12N 1590

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059622490

ABSTRACT:
A method for identifying a cell or strain of cells containing a mutation in a gene involved in growth, comprising the steps of forming a labeled set of strains comprising a plurality of members, each member of the set containing an exogenous DNA fragment of a defined length stably integrated into the chromosome of a member, the defined length in each member differing from the defined length in other members, subjecting the labeled set of strains to mutagenesis so as to obtain mutants from each member of the set of strains, and introducing the mutant strains into a growth environment for a period of time sufficient for growth of a non-mutated strain and determining which strains have reduced growth compared to a non-mutated strain, by determining the presence and size of exogenous DNA fragments relative to each other using PCR and agarose/polyacrylamide gel electrophoresis.

REFERENCES:
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Pike et al., "Development and design of a novel in vivo chamber implant for the analysis of microbial virulence and assessment of antimicrobial therapy," Microbial Pathogenesis 10:443-450 (1991).
Rosey et al., "Dual flaA1 flaB1 Mutant of Serpulina hyodysenteriae Expressing Periplasmic Flagella Is Severely Attenuated in a Murine Model of Swine Dysentery," Infection and Immunity 64:4154-4162 (1996).

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