By David Pike
Texas Woman’s University
October 25, 2022 – DENTON – There is a dimension unknown to all but an unusual few. It is huge, but it fits in the nucleus of a single cell and challenges the concept of the finiteness of the human mind. It is found in every living creature on Earth and within its womb is the blueprint of every organism, but until very recently it existed beyond human understanding. It is the world of the infinitesimal.
It is also the class of Texas Woman’s University biology professor Catalina Pislariu, PhD.
And it’s a little hard to wrap your head around.
“It’s so abstract,” Pislariu said. “When I started studying it at school 22 years ago (when she started doing her PhD) I was at the level of many of my students who can’t wrap their heads around it. How do you do that?
Pislariu, professor Nathaniel Mills, PhD, and teaching assistant Hala Samara teach classes in molecular biology, a branch of science that studies DNA and includes cloning. Pislariou, however, defies this limitation.
“It’s not just cloning, it’s molecular techniques, methods and instruments,” he said.
And it’s one of TWU’s most hands-on graduate-level offerings.
“This is the type of course you can’t teach online,” Pislariou said. “It is intended for incoming graduate students to familiarize themselves with molecular techniques so they can use them in their own research.”
Molecular biology, a phrase that did not exist until 1945, is the study of how molecules interact with each other in living organisms to carry out the functions of life and is applicable to a number of scientific fields.
“In our class we had cancer biologists, neuroscientists, people with protein degradation,” said student Miles Gladen. “We all bring our research to the classroom.”
But something amazing happened in the 2021-22 class. The students didn’t just learn how to make discoveries later in their careers. They actually made discoveries about many unique DNA sequences that were published in the National Center for Biotechnology Information gene sequence database at the National Library of Medicine.
“At first, I didn’t know that the students would get data of such high quality that it could be published,” Pislariou said. “They’re doing every little bit of work that a seasoned scientist would do in their research. It’s a wonderful experience. They went from identifying a gene to moving that gene onto a plasmid, sequencing it, figuring out the sequence and coding, locating the protein at work. of a semester”.
And the National Center for Biotechnology has verified the discoveries as new and correct.
“They were excited,” Pislariou said of her students. “The excitement that brings when you have a project that has a purpose, that starts with an initial question and ends with a final product that gives you a scientific answer, makes them feel really excited.”
For the vast majority of the population, molecular biology is completely foreign. Most of us are ignorant of its concepts and language. For example, consider the title of one of the four published submissions:
“Medicago truncatula eat tricycles Nodule-specific PLAT/LH2 domain protein (NPD2) mRNA, full cds”
Or the title of one of the course research methods from the TWU catalog:
“Plasmids as vectors for recombinant DNA”
And if that’s incomprehensible, try to wrap your head around the physical nature of DNA.
The invisible world of microbes and cells is measured in micrometers. A microbe, such as a bacterium, can be from one to 10 micrometers. An animal cell is 10 micrometers. For comparison, a meter (a little longer than a yard) is one millionth of a micrometer.
But many of us at some point in school looked through a microscope to see germs and cells. DNA, however, resides in a sub-invisible world penetrated by an electron microscope. In fact, it was only 10 years ago that the DNA double helix was first photographed in blurry, indistinct images.
Your cells contain your DNA, made of things called nucleotides, which are 0.6 nanometers. A nanometer is 1,000 times smaller than a micrometer, or one billion times smaller than a meter. However, these tiny organic molecules are the building blocks of the 3.2 billion base pairs of your DNA contained in each of your cells.
If that doesn’t sharpen your perception of reality, try this: if the DNA from a single cell were unwound, it would be six feet long. And if the DNA from all your cells were unwound and twisted end to end, that strand would be 67 billion miles long – 22 times the distance from Earth to Pluto.
“My friends outside of school and academia say ‘what do you do every day?'” Gladen said. “There’s a very high level of knowledge that students have to bring to the class. It’s not just for anyone to get into. But for people who want to delve into the scientific realm, it’s an important class.”
Why so important?
“We’re scratching the surface of molecular biotechnology, with uses we can’t imagine now,” Pislariou said. “And more and more are coming. I think the future of medicine is going to involve a lot of gene therapy.”
Gene therapy involves the treatment of diseases by repairing or reconstructing defective genetic material, which promises to be of enormous importance in various medical fields.
This potential is reflected in the expected growth of jobs in molecular biology. In the next 10 years, the sector is expected to grow by 19%, much higher than most industries.
The molecular biology course gives students the tools they need to be part of this industry. The class draws students from other schools, including the University of North Texas, and, working in small groups, students go through each step, from creating solutions to using liquid nitrogen to grind tissue before extracting genomic DNA.
“I give each student group a gene identification number,” Pislariou said. “From that, they have to figure out how they can recover the sequence using bioinformatics tools, how to clone that gene into a series of plasmids in a way that will eventually produce the encoded protein as a fusion with a fluorescent marker, a green fluorescent protein through a series of transformations, the expression plasmid is grown in tobacco leaves. Within two days, the fluorescent protein will glow, allowing us to know exactly where within the cell the protein is.”
There’s no practical use for making a tobacco leaf that glows, unless you want to roll your own cigarettes in the dark. The purpose here is to learn the techniques and methods of using tobacco leaves as tools to locate the glowing proteins.
“It was one of the best lessons I had in how to do research techniques,” Gladen said. “Many of the techniques I had heard and been taught in class, but Dr. Pislariou and Dr. Mills were able to guide us and did an amazing job helping us understand how to run them and showing us techniques we might not do every day in our research. It’s a real foundational course for a PhD student.”
“You don’t solve everything in one session,” Pislariou said. “It’s the workflow. They do everything, every little step. Every step has its own benefit. Even if one step doesn’t work well, you learn from it. Sometimes you learn better from a mistake than when everything goes smoothly.
“It takes a lot of patience. And passion.”