A new operon is discovered in cyanobacteria that allows the bacteria to breakdown diesel gasoline. W Show more A new operon is discovered in cyanobacteria that allows the bacteria to breakdown diesel gasoline.
When the diesel is broken down it turns from a clear yellow color into a dark purple liquid with a brown solid precipitate and white floating foam. You know that there are the standard regulatory regions and that there are four structural genes but you want to further characterize this operon. The genomic region containing this operon is approximately 9 kB in length and it was recently discovered that on either end (but not in the middle) of this operon that there are NotIrestriction sites. In order to characterize this operon and for later use you want to clone this operon into E. coli. Describe in detail the steps that you would go through to clone this operon from the cyanobacteria into E. coli. Be sure to give all relevant steps ways to ensure that the cloning was successful and a way to test for functionality of the cloned operon. Discuss a method that you could use to determine the order of the genes in this operon (assume that the F gene produces a brown precipitate the U gene produces a red liquid that the L gene produces a white foam and the E gene produces a blue liquid when exposed to media containing 10% diesel). Fully describe one way that you could determine the sequence of DNA encoding this operon. Compare and contrast this method with one other method that you choose not to use and indicate why you feel your method is the most appropriate. Show less
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