By Luke Jenusaitis
Imagine little space shuttles landing on a large blob and injecting a smart little blue alien man into it. The large blob seems to be unscathed from the outside, but inside something is happening. This little blue alien, with the vast knowledge and information he has collected, is able to combine all of the parts of the blob to make thousands more space shuttles and replicate thousands more little blue aliens. Eventually the blob bursts and these spaceships are free to infect more blobs. Seems like something out of a science fiction story right? Wrong! These space shuttles are like bacteriophages, the little blue alien is the bacteriophage’s DNA and the giant blob is a bacterium. For the past two months we in the Johns Hopkins Phage Hunting class have been attempting to find, isolate, and purify the DNA of a bacteriophage that attacks the bacteria M. smegmatis.
The way Dr. Schildbach and Dr. Fisher (Doc Fish) talked about the process, I thought it would be a breeze. Oh how wrong I was. After six Direct Platings and an Enrichment plating I found a couple putative plaques but in the end all failed. I decided to adopt the phage my partner Ellen Bruner was working with. Her phage seemed to be working very well on the plate until I got involved. I thought that perhaps I just brought horrible luck into the equation when we both plated Ellen’s phage and nothing turned up. We attempted to purify the phage from the plate that yielded multiple plaques and again our platings yielded nothing. After the intervention of our wonderful TA, Victoria (aka my BFF, or Vesuvius), we attempted to compare the top agar of Ellen’s plate to the top agar of M. smegmatis. Eventually we learned that the original plate with all the plaques had contaminating bacteria, not M. smegmatis. Essentially what Ellen had found and isolated was a phage that targeted the contaminating bacteria, not the M. smegmatis bacteria. Our happiness disappeared faster than Charlie Sheen’s sanity.
Clearly there was no way I was going to quit. I wanted a phage to call my own. I tried one last Enrichment sample from the Bufano Gardens. From the sample both Ellen and I plated, we found plaques. My top agar moved before it solidified so it looked messy on the Enriched plates, but regardless I still was able to find plaques which was a huge achievement. I could not have been happier. Our next step was to streak the phage at least three times. One of my streaks could not have looked better.
From observing our plaques on the streaked plates we found that they were extremely tiny (about perhaps a 1-2 millimeters) and cloudy, but thank the good Lord they were plaques. Throughout the three streaks we found no difference in morphology. Since we have been taking care of this phage for a couple weeks now we needed to come up with a name. Since I found the phage I wanted to name him John Belushi after my favorite actor. Instead Ellen suggested, Land Shark after the famous skit and because our phage, like the Land Shark was so difficult to capture. We are currently starting our Phage-Titer Assay with Land Shark and will continue on with that process.
Having no microbiology lab experience, I had no clue what to expect. With a small class, two TAs and two professors, we have become a family. We have the quiet ones, the loud ones, the tall ones and the short ones. We have music of all sorts playing from Jimi Hendrix to Broadway show tunes. We have best of friends like “Big Ben” and I. We even have some romance in the lab. Lets just say some phage hunters have become truly infatuated with their phage. The TAs have become our mentors inside and outside of the lab or when I’m just on my way to get a bagel, giving us advice on anything and everything. Just how the hunters have adopted our phages as our children, so too have Dr. Schildbach and Doc Fish adopted all of us as their surrogate children in a way. They made sure to tell us jokes when we were down or just when we were waiting for the phage to infect M. smegmatis.
In the end as long as I have a phage, I’m WINNING.