Louise wrote:I think you are on the correct track (and what you propose is what many researchers are doing). I cannot recall where you are planning to work, or what resources you have, so some of your ideas may be overly ambitious for someone in high school. If you could find a college mentor working in this area, I think you could have a great time. What city are you in? [Don't give me any more detail than that; I just want to search for the research institutions near you.]
cicichen wrote:Louise wrote:I think you are on the correct track (and what you propose is what many researchers are doing). I cannot recall where you are planning to work, or what resources you have, so some of your ideas may be overly ambitious for someone in high school. If you could find a college mentor working in this area, I think you could have a great time. What city are you in? [Don't give me any more detail than that; I just want to search for the research institutions near you.]
Thank you for asking! I live in the San Francisco bay area (city of Cocord). I don't know whether you are familiar with the Joint Genome Insititute (JGI), it is actually close to where I live. And since my science teacher has a second job there she introduced me a mentor. I talked about my project idea with my mentor because I wasn't so sure if I would be able to do this project. My mentor thinks it is doable, but I need to work hard on it.
cicichen wrote:I talked with my mentor about my experimental design.
Here is what my mentor and I came up with:
My goal is to genetically engineer a type of fermentive yeast that produces ethanol so that it is capable of coverting cellulose to ethanol in a one-step process.
First I need to find out several genes that are able to together break down the structures in biomass and produce cellulase to convert cellulose.
Second I need to run a PCR to duplicate the genes
Then I will need to ligate the genes and do a knock in to insert the genes into the genome.
I will have two sets of yeast: the control yeast (no genetically engineering) and the yeast that have been inserted the cellulase produced genes.
I will feed both types of yeast with glucose source, then after a while I will change their food source to cellulose (maybe a form of liquid).
The objective is to see whether the genetically engineered yeast will live with the cellulose source, if it does, then it proves that the yeast can breakdow the cellulose (which is something they are not supposed to do)
My second objective is to measure the ethanol produce as bioproducts from the yeast and to determine its efficiency.
My third objective is to see how the genetic mutation (using UV light) affects the yeast and its ethanol productio.
cicichen wrote:Saccharomyces cerevisiae produces several Î²-1,3-glucanases, but lacks the multicomponent cellulase complexes that hydrolyse the -1,4-linked glucose polymers present in cellulose-rich biomass as well as in haze-forming glucans in certain wines and beers.
cicichen wrote:We have introduced into S. cerevisiae a functional cellulase complex for efficient cellulose degradation by cloning the Endomyces fibuliger cellobiase (BGL1) gene and co-expressing it with the Butyrivibrio fibrisolvens endo--1,4-glucanase (END1), the Phanerochaete chrysosporium cellobiohydrolase (CBH1) and the Ruminococcus flavefaciens cellodextrinase (CEL1) gene constructs in this yeast.
cicichen wrote:The END1, CBH1 and CEL1 genes were inserted into yeast expression/secretion cassettes. Expression of END1, CBH1 and CEL1 was directed by the promoter sequences derived from the alcohol dehydrogenase II (ADH2), the phosphoglycerate kinase I (PKG1) and the alcohol dehydrogenase I (ADH1) genes, respectively.
cicichen wrote:In contrast, BGL1 was expressed under the control of its native promoter. Secretion of End1p and Cel1p was directed by the signal sequence of the yeast mating pheromone -factor (MF1), whereas Cbh1p and Bgl1p were secreted using their authentic leader peptides. The construction of a fur1 ura3 S. cerevisiae strain allowed for the autoselection of this multicopy URA3-based plasmid in rich medium. S. cerevisiae transformants secreting biologically active endo--1,4-glucanase, cellobiohydrolase, cellodextrinase and cellobiase were able to degrade various substrates including carboxymethylcellulose, hydroxyethylcellulose, laminarin, barley glucan, cellobiose, polypectate, birchwood xylan and methyl--d-glucopyranoside. This study could lead to the development of industrial strains of S. cerevisiae capable of converting cellulose in a one-step process into commercially important commodities.
cicichen wrote:I would really like to know what those genes: BGL1, END1, CBH1, and CEL1 do and how do they work together?
And what's a Î²-1 4-glucose?
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