Protein engineering techniques

beachbum · Post by **beachbum** » Wed Jul 05, 2006 1:33 pm

Hello, I am interested in conducting a project where protein (protease specifically) engineering would be required. I am familiar with directed evolution, however, what other techniques are out there?

I've tried NCBI, GoogleScholar, Google and pretty much every search but only very broad information is provided. Some links/literature would be greatly appreciated.

carolinethorn · Post by **carolinethorn** » Wed Jul 05, 2006 1:53 pm

Hi,

It would be useful if you could give us some more information on the aim of your experiment. The types of protein engineering I am thinking that you might want to consider are dependent on what you are trying to change about your protein of interest.
Most of the techniques require that you have made a vector containing the DNA sequence that makes the protein you are interested in and a DNA element that causes the protein to be made when you put the vector into cells (either bacteria, yeast or animal cells)
A commonly used technique is site-directed mutagenesis. This is a way to change one amino acid in the protein at a time.
Another way is making a fusion protein - taking part or all of one protein and joining it to another.
Both of these require manipulating the DNA so would be quite a complex project.

Post back with your plans or ideas and we will try and help out,
Caroline

beachbum · Post by **beachbum** » Wed Jul 05, 2006 2:34 pm

Thanks for the reply. The aim of my project is to engineer a protease for proteolysis of prions. I will most likely be using computational techniques to determine the structure of the engineered protease but I currently have in mind using directed evolution. I am not sure how to do the rest of it.