Science Buddies: "Ask an Expert"

My partner and I are trying to transform the E. coli bacterias protein into a whole new protein by adding a plasmid so it can turn into or something similar to the protein pepsin so it may cure or help with Cystic Fibrosis. The problem is we do not know what specific plasmid can be useful for this or if there is any.

Hi,

This is a really original project idea that would involve cloning. This is definitely a very challenging project. The following is some information that should help you get started.

Here is a project from the Science Buddies website on cloning:

https://www.sciencebuddies.org/science- ... p024.shtml

Here is a link for a site that includes the DNA sequence for human pepsin.

http://www.uniprot.org/uniprot/P0DJD7

Here is an abstract that reports the expression of porcine pepsinogen (the precursor to pepsin). Unfortunately, the complete article is not available.

http://www.ncbi.nlm.nih.gov/pubmed/12135554

Here is a similar article that includes all of the details of the materials and methods. This paper should be useful to you.

http://www.jbc.org/content/272/30/18855.full

I recommend doing additional internet searches for “recombinant pepsin (or pepsinogen),” and related topics to look for additional details on the procedure you would be using for this project.

Do you have access to a laboratory? Do you have experience in using pipettes and sterile technique?

Please post again if you have more questions.


Donna Hardy