Date of Award

5-2014

Degree Type

Honors College Thesis

Department

Biological Sciences

First Advisor

Hao Xu, Ph.D.

Advisor Department

Biological Sciences

Abstract

Mast cells play an important role in the immune system by releasing chemicals such as chemokines and cytokines once they are stimulated. These products are released after stimulation by a process called mast cell degranulation. Mast cell degranulation is accomplished when vesicles containing the chemicals inside the mast cell fuse with the mast cell membrane via SNARE-mediated (Soluble NSF Attachment Protein Receptors) membrane fusion. This family of proteins consists of syntaxin, SNAP 25-like protein, and synaptobrevin/VAMP (Vesicle Associated Membrane Protein)(2). Comlexin isoforms (complexin 1,2,3,and 4) have been known to regulate this system in a fashion that is still unclear. In order to study the mechanism in which these complexins regulate SNARE-mediated membrane fusion, each isoform was cloned and ligated to the pTYB12 vector to be expressed in E. coli. An induction process using IPTG was used in order to induce production of each isoform via the T7 promoter. In this experiment, we were able to clone all of the complexin isoforms, but only complexin 1 and 3 were successfully expressed.

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