The green fluorescent protein (GFP) is a protein, comprised of 238 amino acids (26,9 kDa), from the jellyfish Aequorea victoria that fluoresces green when exposed to blue light. It is a powerful method for nondestructive in situ monitoring, since expression of green fluorescence does not require any substrate addition .
E. coli is frequently studied in microbiology and is the current “workhorse” in molecular biology. Its structure is clear, and it makes for an excellent target for novice, intermediate, and advanced students of the life sciences.
Error-prone PCR in realistic reaction conditions is predicted to be less effective at generating grossly divergent sequences than the original model. The estimate of mutation rate per cycle by sampling sequences from an in vitro PCR experiment is correspondingly affected by the choice of model and parameters. Random mutagenesis constitutes an important approach for identifying critical regions of proteins, studying structure-function relations and developing novel proteins with desired properties.

I’m working on a research project since August 2007, entitled “Generation and Characterization of GFP variants using Error Prone PCR and Characterization in Escherichia coli”. My mentor is Dr. Michael Rubin. I selected this area because after my genetics class with Dr. Rubin I was very interested in the field of genetics. I’m having a great laboratory experience thanks to my group that is composed by my mentor Dr. Rubin and my laboratory partners Erick Aponte and Angiemar Maldonado. This experience has been great because everyday we do something new in the lab that is interesting to learn about. I participate at least once a week meeting with my mentor and my laboratory partners to discuss the work of the week and plan the meeting times for the experiments. We discuss the theory and techniques of each experiment. My team group is formed of responsible, intelligent, and friendly collaborators making our work experience in the lab comforting and high-quality. We have one goal and that is our research – it keep us together and I really like the maturity and commitment of all the people that are part of our research team.

I have learned many techniques beginning with bioinformatics analyses including downloading DNA and protein sequences and comparing them to databases. I have used these sequences to design primers for PCR amplification. Working in the lab I have learned how to carry out many techniques in molecular biology including: setting up PCR amplification reactions, purifying PCR products, separating PCR products using agarose gel electrophoresis, cloning DNA fragments using vector ligations and bacterial transformation.

Our research team has established as a specific aim for this semester to work with the products obtained during our work last semester. New GFP variants synthesized using Error Prone PCR will be analyzed. The first thing we are going to do is to send the mutated fluorescent GFP products for sequence determination followed by DNA analysis. These cloned DNAs will be transformed into the salt tolerant extremophile Halobacterium.

If you want more information related to my research you can click here

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