(if you’re here for the first time look at Blogs 1 -15 in earlier posts )
Maddie mulled over the Black Caucus meeting while driving to Arlington Experiment Station. It was a cold but sunny morning; the crisp blue sky displayed against the white snowy countryside. She also thought about the new equipment she had convinced Dr. Gates to invest in, and how much easier it had made her work. It consisted of a gel apparatus which was miniaturized and already assembled in such a manner that the gel solution could simply be poured into it. No more tedious assembling of glass plates held together with clamps and Vaseline. Those were always leaking and took a long time to run. She couldn’t imagine why the group had kept on using the antiquated method when new equipment had been around for several years. Now, she could run a gel in under three hours.
Last week she had run another set of electrophoresis gels with the varieties containing band 34, all lined up side by side. She wanted a good photo for her thesis. She had been puzzled, however, by the resulting gel, because the stained bands did not all look identical. Some were thin, others were fat dark blobs, and some of them almost seemed to be blurred; it looked like one line smudged into two.
What does it mean? Could it be simply a lab variation? Had she loaded uneven amounts of protein extract on the gel? But she had been very careful to shake the test tubes and load exactly the same amount on each column. Then, it occurred to her that perhaps the wild varieties themselves had varying amounts of the protein. Or that perhaps there were different forms of the same protein in the wild varieties. This could be true if the gene that produced the protein had undergone several mutations.
What if there was a correlation between degree of resistance and amount of band 34 protein? Or, as could be the case, a correlation between degree of resistance and a specific type of band 34 protein. She began to see this as a door opening another dimension to the exploration of the problem. It became important to pursue. What if there was a variation of band 34 protein that conferred no resistance at all?
When she broached the subject with Dr. Gates, he wanted to see the photos.
“That’s a very good observation, Maddie; if I had looked at these gels more closely in the first place I could have been able to tell you there was some suspicious variation there.”
“Do you think it’s possible that we have some allelic variants here?” Maddie asked.
“Well, it’s not possible to detect that by the eyeball method. What you should do is run a two dimensional gel.”
“What does that do?”
“Well, that will separate the protein into its subunits. If these proteins separate into different subunits then you’ll know they are variations of the same protein.”
“Mutations of the same gene?”
“Yes, it’s not too far-fetched. After all, the variation in the wild environment could have easily induced the development of different gene forms.”
Maddie was getting highly charged. She had an intuitive sense of the elasticity of nature in the fight for survival, both on the part of the plant and on the part of the pathogen. If several genes had arisen in the wild, in response to the highly changing pathogen, what was the relationship between them? Which form had arisen first, which second? She saw the potential to solve a puzzle which might help to better understand the mechanism of mutation. Even though her thesis problem was a highly applied one, dealing with food production, it was theoretically possible that any bit of knowledge gained in that work could have wider application, to other fields, other sciences. People were still trying to understand how the AIDS virus mutated so rapidly, how cancer cells developed. This was exciting!
“How do I go about learning the technique for two-dimensional protein separation?” she asked Dr. Gates.
He thought for a moment. “Well, the last one in our group to use it was Jay Wilson. He’s been gone for several years. Why don’t you look over his thesis for the procedure. Aside from that, I think you’re on your own. I would advise searching the literature for modifications.”
Maddie was a bit miffed. Learning a whole new technique on her own was not going to be easy. Often, the literature articles failed to mention bugs in the system and possible pitfalls and one only discovered these from doing the procedure the wrong way several times over. But then, that’s the glamour of science, she reminded herself.
There was one thing she was thoroughly proud of though, and that was the fact that she had made crosses with all the band 34 lines! If she had only used one or two and discarded the rest she would have to go back now and begin those crosses. Whatever the variation turned out to be now, she had at least created lines with all the different genes. The progeny of those eight crosses was now growing in the greenhouses at Arlington Experiment Station.
Early on in the winter semester, Maddie had obtained space in the greenhouses at Arlington to increase the seed from her crosses, so that there would be enough seed to grow her experiments in summer. The work of setting up the planting took several days of soil mixing and dragging wheelbarrow after wheelbarrow of soil into the greenhouses to fill over a hundred pots. After planting, the plants had progressed fairly well, with only an occasional visit needed to check up on things until she began another round of backcrosses to the cultivated parent. Arlington was only an hour from Madison and so these trips were more manageable than the ones to Hancock Station.
She still shuddered to remember the harvest last fall. It had been snowing already when they pulled the last plants out of the field, and bitter cold weather for the subsequent threshing in the unheated barn. Dr. Gates’ graduate students had worked long days under Edgar’s supervision, wearing their gloves and coats all day long. Each harvested row had been placed in a burlap sack, and then dried in the heated seed drying sheds. They had then carted the bags to the barn and fed them through the thresher one by one. She could still smell the dirt that covered the burlap bags and taste the dust that blew out of the thresher. At night, when she came home, she would stand in the hot shower for an hour, just trying to get the grime out of her hair and empty her nostrils of the black residue.
The plants at Arlington were now beginning to flower, and today she would start making crosses. She pulled up on the brittle snow near the frozen apple orchard and parked her car. She entered the greenhouse complex through the general warehouse, where all the soil and equipment was kept and proceeded down the corridor to the Bean Project greenhouses, opening and closing greenhouse doors behind her as she passed through. Despite the cold outside the sun was beating down through the whitewashed glass ceiling making the greenhouses warm and steamy. She started shedding her coat and gloves as she walked through. At one point she looked out of the window to the adjacent row of greenhouses and stopped cold, hand on a doorknob. Working in the facing greenhouse was a magnificent figure of a man, in faded blue jeans and naked to the waist.
Without surprise she recognized the figure of John Pitts. As he bent and lifted the bench boards or the concrete bricks beneath them, in an apparent reorganization or cleaning effort, his Herculean movements left her momentarily agog. Maddie swallowed. What passed through her mind in that moment was sheer admiration for the beauty of the human male. How a man could put forth such physical exertion without complaint somehow emphasized the difference between the male and female graces.
Quite suddenly, Maddie realized that she could be caught peeping, and ripped her gaze away. Girl, what’s the matter with you! Go get to work!
As harshly as one side of her mind chastised her, the other side was laughing with some kind of giddiness that made her lightheaded. Images were hard to erase. But the harsh side won out and by the middle of the afternoon when she ran into John at the water fountain, this time with his shirt on, she barely acknowledged his friendly hello, worried that any revealing vestige of attraction might remain on her face.
She was unaware of her non-verbal cues, or that she could be giving off the impression that she disliked him. So she didn’t notice the frown on his face or the shrug that indicated he was done trying to bridge the gap between them.
That attitude still lingered two weeks later when Maddie and Lisa walked into Pasqual’s Southwestern Deli, where a celebration dinner was taking place in John’s honor. He had just passed his prelims that afternoon.
“John,” Lisa hailed him loudly as they approached the table, “I heard you kicked some butt today.” He got up as they approached and she reached up to slap him on the back and then hug him. Maddie was always intrigued by the fact that Lisa treated John like a big brother.
“Nah, it wasn’t exactly like that,” he said, still holding the petite girl and smiling down into her face. “I just answered their questions. They were okay.”
Maddie murmured her congratulations, which he accepted matter-of-factly. He then asked the waitress for another pitcher of beer while Maddie quietly ordered a coke and put in a request for her food.
They had put several tables together and Maddie acknowledged several other familiar faces; Joel was among them, and Savannah was sitting beside John.
“John, I still can’t believe you asked Dr. Heckley to be on your committee,” she said. “That man’s scary! He’s messed up so many people.”
“Eh, I like the old guy. He didn’t really give me a hard time. I thought his questions were fair,” John responded.
“What did he ask you about?”
John went on to recount episodes from the exam. As she reached for the chips and salsa, Maddie realized she had forgotten to eat all day and that, as usual, she was feeling strung out. When her blue corn cheese enchiladas arrived, with the guacamole, sour cream, black beans and rice on the side she attacked them, reveling in the melted cheese. After a couple of mouthfuls she felt herself becoming human again. She looked up across the table to find John’s smiling eyes on her. She swallowed hard and turned to the person next to her to make conversation.
For the rest of the evening John gave Savannah his undivided attention, never looking back in Maddie’s direction. Maddie ate her food and thought about Savannah’s blood red fingernails, which came to rest so frequently on John’s biceps. She wondered if the woman slept in her make-up. She had never seen her a day without it.
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QUESTION 16: Talk about science, physical attraction, frustrations of graduate school. Whatever.
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