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Operon Fusion

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Abstract

The disruption of a gene pathway was assessed using operon fusion to determine mutation by a bacteriophage, Mu. Two bacteriophages similar to Mu were used, MuD1 and MuCts. The pathway used to determine insertion was the arabinose pathway. A strain of E. Coli was used called MC4100. This strain of E. Coli was infected and diluted a number of times, then plated on 4 different kinds of plates (LBC, LacMac+Amp, AraMac+Amp and AraMac–Amp). A phage control and a cell control plate were also created. Colonies were assessed by color, which determined genotype.

The mutation frequency of Ara+ and Arar mutants were found based on colony forming units per milliliter plated. The mutation frequency of Arar: 2.5E-8. The mutation frequency for Ara+: 1.0E-7. In our second set of data, the mutation frequency of Arar: 6.0E-8. mutation frequency of Ara+: 2.0E-8 based on supposed data given to us by the instructor. The red colonies on plates 10-1, 10-2 were outnumbered by the white colonies except on a 10-3  dilution on Lactose MacConkey + Ampicillin plates where the red colonies were more numerous than the white.


Steric Hinderance - Sec 002 - BIOL 302L - FA09