Researchers names and affiliations Research Title NSF HBCU-UP Program Fort Valley State University Researchers names and affiliations Glass Bead Transformation Results Abstract More than 7k are deleted from A10 genome causing 3 genes to be impacted. We extracted each gene from a bacterial artificial chromosome (BAC) vector and transformed them individually back into the mutant in order to restore the fatty acid phenotype. We then used gas chromatography in order to see if the gene complemented the mutant. Genes one and two where transformed back into the A10 mutant using the glass bead transformation technique The cells and DNA were vortexed for 15 seconds in a glass tube which contained 0.3 g of 0.4 mm glass beads. The cells were then poured in tris acetate phosphate plates that contained the antibiotics hygromycin and ampicillin. 5 days after transformation. Part of BAC plasmid with Restriction Sites AvrII MfeI BamHI Gene #1 Gene #2 Gene #3 1 day after transformation Gene #1 Unknown Conserved Function Only partially deleted Gene #1 was extracted using a single Digestion with enzyme AvrII Gene #2 Codes for an RNA-binding protien Completely deleted Gene # 2 was extracted using a double and single digestion with enzyme MfeI and enzymes MfeI paired with BamHI Gene #3 Partially deleted Only the 3’ UTR is affected. For the #1 gene 42 putative transformants were tested and 10 were found to be positive co-transformants 1-1 1-4 1-5 1-7 1-8 1-10 1-20 1-23 2-17 2-20 2-35 C AT Wt Deleted Problems and Setbacks Contamination Introduction #1 gene from AVRII single digestion Triacylglycerol (TAG) WT Chlamydomonas Reinhardtii Non-polar lipid Highly reduced form of carbon for storing energy Capable of containing longer carbon chains than gasoline Since we were unable to isolate the #3 gene we speculate that some random mutation may have occurred in the back vector which is why our enzymes didn’t work. The BAC vector is currently being sequenced. DELETED About A10 mutant A10 has a low oil phenotype compared with mutants and wild type. And a lower growth rate than wild type. MfeI and BamHI double digestion. MfeI single digestion Conclusion Deleted Four different double digestions were used to isolate the gene. No digestion could successfully isolate the gene. Acknowledgements Future Research