Q. How do you deal with discrepancies between mRNA and protein expression?
How do you deal with discrepancies between mRNA and protein expression?
In life sciencies the expression of specific markers can be measured on the level of proteins (ELISA, western blot etc) and on the level of mRNA (qRT-PCR, micro-array). However, often it is found that the expression levels of mRNA and protein do not correlate. How should scientists deal with this?
1 Answer
Hi,
there are a variety of ways this can be done, but it depends on the organism you study. In most prokaryotes for example, one could make a translational fusion between your gene of interest and an enzyme/tag, which you can integrate locally into the chromosome. this way, the endogenous promoter will drive the expression of your gene (which you can measure with qRT-PCR) and protein (enzymatic assays).
to some degree, this is possible in some eukaryotes as well, provided that they can be transformed through homologous recombination. If not, the only alternative is to measure gene expression (mRNA-SEQ) and do proteomics at the same time. these will only give you a measure of correlation though.
in the end, protein levels are most important. this is what i would focus on given the choice
answered by huitemae