In the past month Dr. Wier and I have continued our research. We have sorted through our potential bacterial candidates and selected a Pseudoaltermonas strain as our focus. Pseudoaltermonas is the genus of a marine bacteria that are frequently associated with marine invertebrates. Many Pseudoalteromonas species produce pigments as well as antimicrobial compounds. We are also using a sample we are calling, “RO7.” Under a microscope I was able to observe the morphologies of the two samples as well as do gram staining. The Pseudoaltermonas sample showed to have rod morphology while our RO7 showed to be coccoid or sphere like morphology.
We recently conducted another polymerase chain reaction (PCR), which as stated in our previous post replicates and amplifies the DNA of our samples. However this time did a PCR and ran a gel electrophoresis. Gel electrophoresis is used in order to separate DNA fragments according to size. Our samples are loaded into small wells of an agarose gel. Then the gel in placed in a buffer in a electrophoresis chamber which allows current to move through the gel. As current moves through the gel, because DNA is negatively charged, DNA fragments move down the gel towards the positive end of the chamber. We will send out our amplified DNA to be sequenced in order to confirm the species of the bacteria that we are using. Our PCR was successful so we are now ready to move ahead. Next steps include making a specific media to grow our samples on in order to move forward with our protocol.
Our next step involves doing a transposon mutagenesis. We are going to take the Pseudoalteromoas strain and introduce random breaks in the genes throughout the genome in order to find genes involved in the pigments involved with antimicrobial compounds. The next stem will involve selecting many mutants and archiving them as well as screening them for different phenotypes.