Discussing results with graduate student Jason Kempton, whose job is to refine 

 field application of the gene probe to study Pfiesteria's geographic distribution 



stars being obscured by a bright moon 

 and city lights. 



Also, breaking into the cells with 

 the probe may prove difficult, Rublee 

 says, because by nature the cyst resists 

 intruders. 



But the challenge is irresistible to 

 this scientist, and he hopes to develop a 

 probe that will penetrate Pfiesteria's 

 cyst form in the sediments, saving 

 precious time and uncovering the 

 organism before it can kill fish. 



"We've been able to amplify 

 [replicate DNA segments] from cysts in 

 the lab, so we know we can break into 

 the cyst cells. Whether we can on site, 

 I'm skeptical," Rublee says. 



Extracting DNA from cysts in the 

 sediments and then probing it is another 

 possibility. One of the first two 

 methods described above - using the 

 probe as a primer or blotting - could 

 then be applied to test for binding. 

 Rublee's graduate student Eric Schaefer 

 is working on a method to extract 

 Pfiesteria DNA from sediments. 



Investigative Teamwork 



Rublee and his team have 

 developed eight oligonucleotide 

 probes that work to varying degrees on 

 Pfiesteria in the laboratory. This 

 success has taken years of patience, 

 perseverance and — as the researcher 

 selflessly points out — the input of 

 countless other people. 



For Rublee, an ecologist by 

 training, the opportunity to work on 

 Pfiesteria arose through his associa- 

 tion with JoAnn Burkholder, the North 

 Carolina State University aquatic 

 botanist who first identified it in 1991 . 

 The two were working together on a 

 different project at the time, but 

 Rublee recognized the value a gene 

 probe would have on the newfound 

 dinoflagellate. The two researchers got 

 funding from North Carolina Sea 

 Grant and the Water Resources 

 Research Institute and have worked 

 since then to solve some of the 

 organism's myriad mysteries. 



But this official collaboration is 



not the only way scientists like Rublee 

 pick one another's brains. He takes 

 questions about molecular biology to 

 members of his department, who then 

 help him brainstorm and troubleshoot 

 as they sip coffee or stroll down the 

 hallways. And Rublee takes advantage 

 of scientific meetings and the ease of 

 electronic mail to consult with a wider 

 base of knowledgeable folks, some of 

 whom may impart that one bit of 

 information that makes the proverbial 

 light bulb flash in Rublee's head. 



"A lot of informal communication 

 transforms into real science," he says. 

 "Science is a community effort, and I 

 really think that the science that works 

 best is that way." 



Sometimes the contribution of 

 other scientists raises more questions 

 than it answers. Rublee recently 

 consulted an expert in phylogeny 

 about whether his assessment of where 

 Pfiesteria sits on the evolutionary tree 

 was accurate. The phylogeny expert 

 concurred with Rublee's suspicion 



8 MAY/JUNE 1997 



