For our research project we are studying marine biofilm formation. We are going to use forward genetics with transposon mutagenesis to attempt to disrupt the genes that code for biofilm formation in an attempt to better understand biofilm expression in the marine environment.
During the past month Dr. Wier and I have been trying to isolate marine bacteria that produce an easily recognizable growth pattern. The criteria include strong biofilm production and rapid colony growth. Numerous samples were taken from the local Orange Striped Anemone as well as Hawaiian water stock cultures stored in Dr. Wier’s lab. Each sample taken was inoculated in a nutrient media tube and placed in a room temperature incubator for 24-48 hours. If after this time a ring like film developed on the top of the sample in the test tube the sample was then streaked on petri plate to ensure only one bacterium was present and could be isolated.
After determining roughly 22 good bacterial candidates from our samples we then performed a polymerase chain reaction (PCR), which ultimately amplifies and copies a small piece of DNA that is used to identify our bacterial samples. The next step was to send out our DNA samples to be sequenced. When the sequences were returned to us we then used the Basic Local Alignment Search Tool (BLAST) to determine the identity of each of out isolated biofilm producing bacteria. Next steps include choosing a few appropriate candidates for the transposon mutagenesis. Now that we have a great phenotype we can break the genes in the bacterium and figure out which ones were involved in creating the biofilm.