Biology assignment primer design
You can draw an image by hand and take a photo to insert into your document if you wish. Only 100 words should be sufficient for each answer, but a max of 200 will be accepted.
Below is part of a hypothetical coding sequence of a eukaryotic gene showing it contains a phosphate binding loop or “walker A motif (G-X(4)-GK-{TS}). The Walker-A motif is part of a universal binding pocket found in a vast array of proteins that span all kingdoms of life. Refer to the attached review articles as a starting point, but include other references you may find.
Your role is to generate an inactive mutant of the protein encoded by the gene sequence below in an E. coli expression system, show the primers you would design to generate that mutant. Base your design on the mutations tested in adenylate kinase reported in Saraste et. al, (1990) (4 marks).
List the important features of the primer that you have checked for to ensure the success of your experiment (2 marks).
Once you have designed your primers, draw out a flow diagram, with a few explanatory notes, showing the experimental steps needed to perform this mutagenesis experiment (refer to the lecture notes for help) (2marks).
