(1) You have discovered an alien species and are sequencing its genome. You find
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Question
(1) You have discovered an alien species and are sequencing its genome. You find a gene that has mild homology to several genes in other species. Some of these genes are thought to be involved in the development of the eye. One is thought to play a role in the developing brain. Another has no known function. You want to see in which mouse tissues the RNA for this gene is expressed (and compare the relative levels from tissue to tissue).
(a) Which of the following would be the best technique to use to approach this problem: Southern blot, Northern blot, in situ hybridization, immunohistochemistry, Western blot?
(b) Explain your choice from part (a).
(2) You look more closely at the sequence of the gene from Question #1, and you find evidence that it is a kinase (an enzyme that phosphorylates other proteins). You therefore want to try to produce large amounts of this protein so that you can do in vitro biochemical experiments with it.
What specialized kind of vector do you need to put the DNA coding for this protein into in order to make the protein in cells?
What would be a good host to try if your goal is to make large quantities of your protein?
What are some possible problems that might occur using this host?
Explanation / Answer
Ans-1-A the best method to detect the expression of RNA in a mouse will be in situ hybridisation.
Ans-1-B in situ hybridisation is a best choice as all other methods are in vitro methods while the in situ hybridisation is a in vivo method so it can easily diagnose expression of RNA inside the living mouse.
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