2022 Student Research Conference:
35th Annual Student Research Conference

Investigating the True Open Structure of the CFTR Protein Using Site-Directed PCR Mutagenesis


Elizabeth J. Thompson
Dr. Stephen Hudman and Dr. Yohei Norimatsu (A.T. Still University ), Faculty Mentors

The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is an ion channel protein encoded by the CFTR gene that allows chloride ions to pass through the cell membrane. Cystic fibrosis may occur if enough functioning CFTR channels are not expressed in the cell membrane, as the flow of chloride ions is crucial in proper hydration of mucus on the airway surface. High-conductance high-open probability CFTR would allow more chloride ions to pass through the membrane. This project’s goal is to mutate the CFTR gene to form a true open structure of the protein as a potential treatment for cystic fibrosis. PCR mutagenesis is performed, and the DNA is circularized, transformed, and isolated. The process is repeated to create another mutation, and the purified DNA is sequenced and verified. It will be injected into Xenopus laevis oocytes, and the conductance will be measured. Increased conductance would indicate potential true open CFTR channels.

Keywords: CFTR, Cystic fibrosis, PCR, Mutagenesis, Oocytes

Topic(s):Biochemistry and Molecular Biology
Biology

Presentation Type: Poster Presentation

Session: 2-18
Location: SUB Activities Room
Time: 3:00

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