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 repressed and where operator sequence would be 4) Describe an experiment to phenotypically confirm how this pathway is regulated using a SELECTION involving styrene catabolism 5) Describe an experiment to phenotypically confirm how this pathway is regulated using a SCREEN involving light production

Answer 1

Ans-1 the following experiment will help in understanding the pathway.

10-20 sets of the bacteria should be plated on the petriplates and they should be observed in such a manner that the catabolism of the sugar should be on individual step in each petriplates and then their composition should be checked by using various instrumental methods.

Ans-2 this type of catabolism can be moved in the E.coli by causing mutation in the responsible gene or by making transgenic E.coli with the help of genetic engineering.

Ans-3 the repression of this pathway will be due to presence of a repressor gene which will change the orientation of the various other genes. The operator gene will be located between the promoter and functional genes.

Ans-4 this can be performed by using stryene operon system . In this a sequence may be prepared which will consist of various genes like repressor, operator, promotor and functional genes. After preparation this operon should be checked for it's proper working.

Ans-5 the same can be checked by using some operon but here we will tag the genes with the fluoroscent tags so when these genes will be expressed they will show the production of fluoroscence light.