You are interested in studying protein interactions with your favorite protein (
ID: 57409 • Letter: Y
Question
You are interested in studying protein interactions with your favorite protein (YFP). Specifically, you wish to determine if YFP interacts with another interesting protein (AIP) in mouse fibroblast cells. Here's what you have to work with: cDNA for the gene encoding YPF cloned into a bacterial vector that has a bacterial promoter a vector that has an enhancer and promoter allowing expression in mouse fibroblast cells antibody against AIP. but only enough for Western blot analysis mouse fibroblast cells that constitutively express AIP Your advisor is asking for the data in one month, not enough time to generate antibody against YFP. Is this experiment possible or is your advisor crazy? Outline the experiments that you would do to determine if YFP interacts with AIP in mouse fibroblast cells. What would your expected results be if they do interact? Briefly describe the different steps and techniques involved. Do not describe detailed protocols but key steps, expected results and verification methods at each step and the final results if YFP and AIP interact. Some here's some additional reagents primers for site directed mutagenesis matrix for affinity chromatography containing antibody against FLAG epitope tag Some more hints add an epitope tag to the cDNA clone the tagged cDNA into the mammalian expression vector perform affinity chromatography using a matrix bound with antibody to FLAGExplanation / Answer
coloning vector is simply known as piece of DNA, which disserated from plasmid (or) virus (or) organism.
which the forgein DNA fragement can be inserted for coloning purposes.
These vectors are having some important features like
a) Convenient insertion
b) Removal of fragment (DNA) from inner side or outer side of vetcor
coloning vector are of many types but Genetically engineer plasmids is mostly used type.
the mouse fibroblast should be run NCBI blast. the samle used run n this blast then the protein peaks will be blast on screen like graphical representation. by that we can find out the protien. the sequencing of protein is also dislay.
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