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Overexpression of NAR1.2 and LCI1 in Chlamydomonas reinhardtii

Undergraduate #1
Discipline: Biological Sciences
Subcategory: Biochemistry (not Cell and Molecular Biology and Genetics)

Courtney C. Bass - Baltimore City Community College
Co-Author(s): Francis Ayehfor and Stephen M. Miller, UMBC, Baltimore, MD Rose Gbemefu and Amrita Madabushi1 BCCC, Baltimore, MD



Grown under optimal conditions algae have the ability to use free resources (CO2 and sunlight) to grow rapidly as well as produce significant amounts of biomass that can be converted into biofuel to be used as an alternative to fossil fuels. The aim of this project is to improve the growth rate of Chlamydomonas reinhardtii. This green alga is a model organism for lab research because its whole nuclear genome is known, and many molecular genetic tools make it easy to manipulate. We are focusing our efforts on the carbon concentrating mechanism (CCM), which regulates CO2 uptake in the organism, and more specifically on two CO2 transporters, LCI1 and NAR1.2. LCI1 (low CO₂ induced) localizes in the plasma membrane and NAR 1.2 functions in the chloroplast envelope. Our overall goal is to overexpress these proteins by ligating the coding sequences for the LCI1 and NAR1.2 genes into C. reinhardtii nuclear expression vector pARG. reinhardtii but a much better commercial production organismand then transforming into C .reinhardtii. Thus far we have succeeded in generating the expression vectors for both LCI1 and NAR1.2, and have obtained transformants for the NAR1.2 vector. Multiple lines of the transformed algae were cultured and are being tested by western blot analysis. We will do growth curve and biomass accumulation analyses on the best expressing strains. If we are successful in improving algal growth by overexpressing NAR1.2 and/or LCI1, the next step will be to express these enzymes in the Chlorella vulgaris, a green alga that is related to C. reinhartdtii.

Funder Acknowledgement(s): The results were obtained as part of the Research Experience and Mentoring (REM) program in the Department of Biological Sciences at the University of Maryland Baltimore County. The program is funded by a grant (REM supplement to NSF-EFRI1332344) from the National Science Foundation (NSF) Directorate for Engineering (ENG) Office of Emerging Frontiers in Research and Innovation (EFRI).

Faculty Advisor: Stephen Miller,

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This material is based upon work supported by the National Science Foundation (NSF) under Grant No. DUE-1930047. Any opinions, findings, interpretations, conclusions or recommendations expressed in this material are those of its authors and do not represent the views of the AAAS Board of Directors, the Council of AAAS, AAAS’ membership or the National Science Foundation.

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