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Engineering a GFP Based Screening Method for Tag-fused Gene of Interest Cloning and (Co)Expression Vectors

Student Researcher: Scott Dougherty, Undergraduate Biology & Biochemistry Major

Sponsoring Faculty: Dr. Matthew Junker, Department of Physical Sciences & Dr. Carsten Sanders, Department of Biological Sciences

Scott DoughteryProject Description:  Living organisms contain thousands of different proteins, some of which provide basic structure to cells and tissues, while others take on a more active role by catalyzing chemical reactions that make life possible and allow its propagation. Studying specific proteins and their functions in these processes allows us to better understand life at the molecular level. A useful method of producing a particular protein for these studies is to clone its gene into a bacterial plasmid. The plasmid is then transformed (inserted) into a bacterial cell, which has all the "machinery" needed to read the DNA and produce a desired protein. The "machine" is then "turned on," or said to be induced by the researcher. Once the protein is made, the cell can be lysed (split open) and the protein be separated (purified). This process is not only useful in studying individual proteins and their interactions with others, but also for large-scale manufacture of medically relevant proteins such as insulin.

The aim of this research project is to design two new DNA plasmids (or cloning vectors) that will be useful in the study of protein function and interaction as described above. In addition to being a new tool for molecular biologists & biochemists in general, we will specifically use them (along with two existing plasmids) to perform functionality and binding interaction studies of human proteins inside of the bacteria Escherichia coli. The two human proteins we will study are related to apoptosis (programmed cell death), which is not yet fully understood but is known to play a role in cancers & neurodegenerative disorders.

 

September, 2013