IGEM:UNAM/2009/Notebook/Modeling logbook Claudia/2010/03/09

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Objective: Obtain the E.coli strain trpR mutant

In planning our project, we have designed a biological device that involves a transcriptional response directed by LovTAP, a chiameric blue light photoreceptor that has fused the DNA binding domain of the tryptophan repressor regulator (trpR).

For this reason we have been looking for an Escherichia coli strain mutant in trpR to avoid the cross-talk of the endogenous function of this gene with our system.

After a careful look at the literature, we found that Dr. Charles Yanofsky had the mutant, so I sent him an e-mail, asking him to provide us the mutant.

Finally we received the mutant, after waiting 3 weeks.

The features of the mutant are:

  • Identification number: CY15001
  • Sex(Hfr,F+,F-,or F'): F-

5 Mutations:

  • lamda- (lambda lysogen deletion)
  • IN(rrnD-rrnE)1 (Inverts the region between rrnD and rrnE son rRNAs)
  • rph-1 (RNase PH )
  • tnaA5 (tryptophanase)
  • trpR55 (represor trpR)

Working on the mutant

In order to test if each colony is genetically what we expect (trpR-).

We have to consider that the mutation is a frameshift. So, we are planning to test them with PCR and sequencing the obtained products.