yeast protein kinase research proposal

Description
specific instructions are attached as file
*** IMPORTANT selected protein kinase to research: DBF2
****plagiarism check
– As stated in the guidelines at least 5 research papers must be used to obtain the necessary background on your kinase and support why studies on its mutant forms may be valuable. These 5 references must be directly related to your kinase of interest. In other words, though you can and should include citations for other papers. Shokat’s procedure will not count as one of these 5 essential protein kinase references.
– Please make sure that you can find your assigned gene both in the Shokat kinase database and on SGD
-A reminder that when you look up the DNA sequence for your protein in SGD you will be provided with the sense strand only.
Each student will be assigned a different yeast protein kinase to research and design primers to

mutate the gate-keeper reSidue.

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Aim: briefly state what the point of the proposal is.

Background: explain why you want to mutate your kinase. This should include a description
of processes your kinase is involved in.

Primer Design: assume that you are provided with an appropriate plasmid containing the
ORF of your asSigned kinase. Follow the stegs outlined in this lecture to locate the go 6-
keeper residue in your kinase and find the NA sequence encoding this region. Your de5ign
sec ion must include the following:

I) You will need to decide which amino acid. Ala or Gly. to change the gate-keeper residue
in your protein to. Better to change very hydrophobic residues to Ala and less hydrophobic
residues to Gly ( . , . , : hydrophobichtml). Why Explain.

2) Make a figure to show how you designed your forward and reverse mutageneic primers
(i.e. where they bind to the template as we discussed in class in lecture 2 slide 7).

3) Write your final primers in 5′ to 3′ direction.

4) Calculate and state the Tm of your primers using the formula on slide 8 lecture 2.

Implications: Research and then briefly discuss why it is important to determine the function
of your kinase by observing the effectswhen it cannot function. Any human homologs
Implications in isease. therapeutics. biotechnology etc.

Note-these are guidelines but NOT a rubric. They are an overview of our expectations butane

not exclusively limited to the above.

In addition to content you will be assessed on scientific.
understanding and use of correct terminology and writing style.

Word Limit: 1000 words (excluding any figure legends).

You must include at least 5 references which must be cited
appropriately in the text (usmg the last name (of the firstauthor) et
a .. or number formatland then listed in a complete bibliography at
the end as follows: paper title, year, ALL full author names. Journal
name, volume and pages. At least 5 references must be published
research articles. WebSl es. textbooks, my lectures etc. do not count.
SGD and pubmed are useful sources to ind appropriate aFiicles.

DO NOT plagiarize from any scientific. source or each other. You must
completely para hrase any information you acquire from any
SCIentific source you are NOT allowed to paraphrase another
student’sWork). e Will checlg every report and if you are caught
With plagiarized material dlSClplanl’Y action Will be taken.