Insertion of the botryococcene metabolic pathway into the nuclear genome of Chlamydomonas reinhardtii
Microalgae based biofuels have been acclaimed as an environmental solution to dependence upon fossil fuels. One noteworthy microalgae species, Botryococcus braunii, produces a unique hydrocarbon (botryococcene) that has garnered particular interest. These botryococcene hydrocarbons can be readily converted into petroleum-equivalent fuels using existing hydrocracking and distilling techniques. Unfortunately B.braunii is a very slow growing microalgae and there have been no successful attempts to genetically modify any of its genome, thus rendering it impractical for industrial applications. Our collaborators from Texas A&M University recently identified the genes responsible for botryococcene production. These identified genes have been titled squalene synthase-like genes 1 and 3 (SSL-1 and 3). Together they convert the already indigenous FPPS into botryococcene. Thus the aim of this project is to insert a construct containing both SSL-1 and SSL-3 recombinant genes into the nucleic genome of the model organism Chlamydomonas reinhardtii. By utilizing ribosome-skipping sequences, it is only necessary to generate a single multi-gene construct via Gibson Assembly. The assembled construct is then to be inserted into the nucleic genome of C. reinhardtii via electroporation and any transformed cells will be identified through an anti-algaecide screening process. Establishing a method of inserting the botryococcene biosynthetic pathway into the well studied, and faster growing,C. reinhardtii will then prove essential when researchers attempt to insert the botryococcene metabolic pathway into the a lesser studied (but industry preferred) microalgae strain such as chlorella.
As part of the PGRP program I have been given the opportunity to conduct cutting edge research in the field of microalgae biofuels research. I was also able to work with our collaborators at Texas A&M University for a week. Initially I found myself on a steep learning curve, although with the guidance of my mentor and fellow interns, I developed a much better understanding of the principals at play. From there the project shifted from being quite daunting to something I grew passionate about. Not only am I leaving the Boyce Thompson Institute a better researcher in terms of raw knowledge and methodologies, but also able to meet dozens of researchers. Who together shaped my understanding of the lifestyle/cultural challenges required being a successful researcher. In these past ten weeks the program (and the people I have met) have humbled me many times and taught me more than I could have imagined about the world of research. This newfound wealth of knowledge has cemented my desire to continue conducting research projects at my home university and re-affirmed my ambitions to enroll in graduate school to persue a Ph. D.