Date of Award

5-2013

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Biology

First Advisor

Cuong Q. Diep, Ph.D.

Second Advisor

Robert D. Hinrichsen, Ph.D.

Third Advisor

Robert J. Major, Ph.D.

Abstract

Humans cannot regenerate their kidneys, whereas zebrafish can regrow new kidney tissues after injury. To study this process, we set out to generate a transgenic zebrafish line in which a specific part of the kidney, the proximal convoluted tubule (PCT) is labeled with GFP. A DNA construct carrying the PCT-specific slc20a1a gene promoter was chosen for this process. First, we modified it so that it would carry the Tol2 cassette for transposon-mediated transgenesis. Then, a switchable reporter cassette (attP1-GFP-2A-CreER-attP2) was inserted into the DNA construct so that the slc20a1a promoter would control expression of the GFP reporter. The attP1/attP2 recombination sites flanking the reporter cassette would allow for future exchange of a different cassette, such as a gene that produces a toxic protein, for PCT-specific ablation. Our data suggest that the final DNA construct was successfully generated and it is ready for injection into zebrafish embryos for transgenesis upon sequencing.

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