Discipline: Biological Sciences
Subcategory: Climate Change
Newton Mayaka Bosire - Baltimore City Community College
Co-Author(s): Stephen M. Miller, Department of Biological Sciences, UMBC, MD
The rising cost of oil prices and the threat of global warming due to emission of Carbon dioxide has led to research of other alternative sources of energy including biofuels produced by algae. The algae produce neutral lipids which are converted into biodiesel in large amounts. In this research, we are focusing on increasing the growth rate of the model green alga Chlamydomonas reinhardtii by overexpressing the gene Carbonic Anhydrase 1 (CAH1). C. reinhardtii is a unicellular alga that can be easily genetically modified. Carbonic anhydrase 1 is located in the periplasmic space between the cell wall and plasma membrane and it is induced by low carbon dioxide (CO2) concentrations. The cell has to maintain high CO2 concentrations to optimize carbon fixation by the Calvin cycle and CAH1 is involved in the interconversion of carbon dioxide and inorganic carbon that helps to elevate internal CO2 levels. We generated a nuclear expression vector that contains CAH1 coding sequence flanked by C. reinhardtii 5’ and 3’ regulatory sequences (HSP70A-RBSC3 hybrid 5’ UTR + promoter and RBCS2 3’ UTR) and transformed into C. reinhardtii, but no CAH1 protein accumulated in any of the transformants. To improve CAH1 expression, we synthesized a bleomycin resistance gene fragment (ble) with a 3’ viral 2A peptide sequence and are inserting this fragment just upstream the CAH1 coding region in our CAH1 vector. Bleomycin-resistant transformants should also express CAH1 because ble expression is coupled to expression of the downstream gene (CAH1). Our plasmids will be transformed into C. reinhardtii and the growth rate will be analyzed and protein accumulation determined by Western blot analysis. If this strategy increases growth rate in C. reinhardtii we will apply it to industrial algae such as Chlorella vulgaris for large scale production of biodiesel.Not Submitted
Funder Acknowledgement(s): This program is funded by a grant (REM supplement to NSF-EFRI-1332344) from the National Foundation (NSF) Directorate for Engineering (ENG) Office of Emerging Frontiers in Research and Innovation (EFRI).
Faculty Advisor: Stephen M. Miller, email@example.com
Role: I did all of the experiment but I am currently in the transforming part of the experiment.