I am expected to design an experiment like the one below. Can you help me find my way to design this experiment? (in means of molecular biology techniques)
I am not so familiar with the techniques since I graduated from chemical engineering. Thanks!
You are working on Interleukin-6 signal transduction pathway for inflammation in cell culture system. When you add Interleukin-6 to cells expressing Interleukin-6 receptor, 100 kDa protein is highly phosphorylated on tyrosine residues. Please use all required advanced molecular biology techniques in order to answer the questions.
a) How do you clone gene encoding this protein, show genomic and also cDNA of your gene encoding 100 kDa protein?
You showed that this is new transcription factor and please show this transcription factor activates some of genes.
c) Please show location of this protein in cells before and after activation.
d) How do you show this protein binds to promoter region of gene (Please put control and make sure this protein specifically binds to this region)?
e) How do you show that which base is important for binding to your transcription factor?
f) How do you show other protein binds to your transcription factor activates gene expression?