Theses and Dissertations

Date of Award

7-26-2025

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Biology

First Advisor

Michael W. Persans

Second Advisor

Hudson R. DeYoe

Third Advisor

Megan E. Keniry

Abstract

With the rising demand for alternative fuels, algal-based biofuels are a promising, sustainable source that meets commercial interest while producing lower greenhouse gas emissions compared to fossil fuels. This thesis explored two studies, evaluating microalgae for their future potential in lipid yield for biofuel production.

The first study involved developing a cost-efficient electroporation protocol that allowed for the genetic transformation of Chlamydomonas reinhardtii using the pChlamy_4 vector, which supports future cloning of genes to enhance lipid production. Parameters affecting DNA uptake and integration into algal genomes were studied: the effect of cell density, the settings of the electroporation device, and the gap size of the electroporation cuvettes were tested. A set of optimal parameters was identified that resulted in the maximum number of algae cells transformed by the vector DNA.

The second study involved identifying five-field isolates of algae that were collected in South Texas via barcoding for potential use in biofuel production. Ribosomal DNA primers 23S (chloroplast genome) and ITS2 (EUK) (nuclear genome) were used to amplify target regions that are within the ribosomal RNA genes. Genomic DNA was amplified using these primers, and the resulting PCR products were sequenced. The DNA sequences obtained were then compared to those of known algal species in databases to phylogenetically characterize the unknown algal species.

Comments

Copyright 2025 Michelle T. Rada. All Rights Reserved. https://proquest.com/docview/3254349751

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