You have just isolated a new strain of Gram negative bacteria that can cataboliz

Question

You have just isolated a new strain of Gram negative bacteria that can catabolize the sugar aspartame. Genome comparisons to all other strains don't provide any insight into the identity of this pathway. 1) Describe an experiment to identify the pathway coding for aspartame catabolism 2) Describe an experiment to move this pathway into E. coli 3) Given what we learned in class, describe how this pathway is likely positively regulated and where promoter sequence would be 4) Describe an experiment to phenotypically confirm how this pathway is regulated using a SCREEN involving styrene catabolism 5) Describe an experiment to phenotypically confirm how this pathway is regulated using a SELECTION involving LacZ

Please list relevant strain/plasmid names, or draw/describe your own custom plasmids for these experiments

Explanation / Answer

Q1. An experiment to identify the pathway coding for aspartame catabolism can be done by:

First using knockin and knockout confirm the gene coding for the enzymes of the pathway.

Supply radioactively labeled metabolic substrates; for the pathway and then trace these radioactive substrates as they move and change in the pathway   

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