Question

The figure below describes the main players in the paper β-catenin 5' ZBP1 protein Binds, regulates other mRNAs 1. p-catenin binds the promoter of the ZBP1 gene. A. Describe one way to measure whether β-catenin binding results in increased or decreased levels of ZBP1 mRNA. (3 points) B. Describe one way to measure whether β-catenin binding results in increased or decreased levels of ZBP1 protein. (3 points)

Answer 1

Question A answer

you have to do promoter binding reporter luciferase assay, where you can clone the proter of a ZBP1 promoter in pgl3 vector if beta-catenin will bind the luciferase expression will high.you can compare the result control versus treated Beta-catenin overexpress.

Question B answer

First, we do a knockdown assay of the Be catenin and check the expression of ZBP1. the knockdown result will show the decrease at protein level you can show by western blot against the anti beta-catenin and anti ZBP1.

and you can clone Bcatenine in over expression Vector overexpression of beta-catenin increase the expression of ZBP1 .and you can isolate the protein from the treated versus control of overexpression vector.

And perform western blot againt the b catenine and ZBP1 you will find the high expression at protein level.