EXPERIMENTS WITH GFP: THE ART AND THE SCIENCE

RUTGERS SUMMER SESSION COURSE (3-CREDITS)
FOR GIFTED and TALENTED PRE-COLLEGE STUDENTS
AND 1ST YEAR COLLEGE STUDENTS


THE OVERVIEW: This summer, 2019, Professor Bill Ward begins his 7th year of teaching “Experiments with GFP: The Art and the Science” to gifted and talented youngsters from area middle schools and high schools. Dr. Ward is a world expert on GFP (green-fluorescent protein), having studied this amazing protein for more than 40 years. Students who complete the course receive 3 college credits, transferrable to almost any other college or university. While our course is rigorous, we maintain a student/teacher ratio of 5 to 1, so students get more personal attention than they will in any other college class. [By comparison, I have taught biology lecture courses having enrollments of 200 or more students, all packed into one huge auditorium.]

Our summer session course is open to Rutgers freshman as well as much younger students. More than two hundred students, some as young as 9 or 10, have gotten excellent head-starts in college by taking this course. One particularly bright student took the course at just 8 years of age. Ninety-nine percent of our registrants have earned passing grades.

WHERE AND WHEN: This hands-on biochemistry lab/discussion course is held in a Rutgers biochemistry teaching laboratory on the second floor of Lipman Hall on the Cook Campus of Rutgers University, 76 Lipman Drive, New Brunswick, NJ, 08901. The course is offered twice this summer – there is a morning section (9 am to noon) and one in the afternoon (2 pm to 5 pm). The morning section and the afternoon section both begin on June 24 and end on July 15. (There will be no class on July 4). The two classes, together, will go on an all-day field trip on July 15 (the last class meeting for both sections). Detailed information about the field trip will follow.

REGISTRATION: We need at least 16 students for any course section to run. We reserve the right to cancel a section that is under-enrolled. But, if sections 1 and 2 reach capacity (20 students per section—40 in all), we are willing to add a third morning section—July 17 to August 6. As before, we need at least 16 students to open a third section, if we have even greater demand, we will open a 4th section in the afternoon (2 pm to 5 pm), July 17 through August 6.

WHAT IS GFP? GFP (green-fluorescent protein) is a Nobel Prize winning protein used in almost every area of biotechnology and biomedicine. Its brilliant green fluorescence makes GFP an ideal teaching tool, as every step in a protocol can be followed in real time. Unlike most biochemicals, that cannot be seen, GFP is always visible. Professor Ward, who began his education in biology, now has the longest continuous biochemistry research program on GFP of anyone else in the world. While primarily a biochemistry lab/lecture course, this is one of a very few courses to integrate the arts with the sciences. Don’t worry, you are not graded on your artistic talents, but you will have an opportunity to perform in an optional talent show near the end of the course.

picture of GFP
E. coli colonies expressing GFP

THE SCIENCE: In a fully equipped university teaching laboratory, we begin by extracting recombinant GFP from genetically transformed, non-pathogenic bacteria (E. coli). Safety is stressed in the lab, as each student wears goggles, lab coats, and gloves. Lab exercises with GFP include: high speed centrifuging to concentrate the cells, cell lysis (breaking open the cells), ammonium sulfate precipitation to collect the protein fraction, or, alternatively, three-phase partitioning (a seldom used, but spectacular method for purifying proteins). Following these steps you will do hydrophobic interaction column chromatography, and then perform a final “polishing” step by gel filtration. Throughout the process, we monitor GFP purity by UV-Vis spectrophotometry and fluorometry. You will experiment with ion exchange separations on membrane absorbers, test pH sensitivity of GFP, and estimate the isoelectric point of the protein. Final purity will be judged by high performance size-exclusion liquid chromatography. Every method used in the lab will be carefully explained by the instructors, in small groups of 4 to 5 students per instructor. We will have four trained instructors, so the student/teacher ratio is never higher than five to one. Questions are always welcomed.

In addition to working with GFP, you will do simple colorimetric tests for peroxidase extracted from soybean hulls and from root vegetables like sweet potatoes, white potatoes, and celery roots. The colored product of this reaction (a teal blue solution) will be used to identify BPA (bis-phenol-A), a potentially harmful chemical found in ordinary store receipts. You will be given prototype BPA test kits to take home to your parents and teachers to test whether your own store receipts contain BPA. You will keep notebooks, write a final lab report, take one short quiz a week, and take a final exam the day before our talent show. If departmental toxicologist, Professor Cooper, is available, we will ask him to talk about his research on the development of fish embryos exposed to BPA. After registration is complete, a 40-page course called “Introduction to Chemistry and Biochemistry,” written by Professor Ward, will be emailed to each student who requests the course. Much of what we cover in “Experiments with GFP” is included in this introductory course.

THE ART: Conducting biochemical experiments with the amazingly brilliant green-fluorescent protein is an aesthetic experience all by itself. But, viewing live comb jellies (ctenophores) glowing in the dark, and then later, seeing their 8 rows of fused cilia diffracting daylight into a rainbow of colors is an incredible spectacle. Artistic aspects of the course are not confined to viewing bioluminescence. In your small groups, you will be given an opportunity to design, on paper, an aquarium system suitable for maintaining comb jellies in an artificial sea water environment. Later, you will create beautiful wall hangings directly from dried comb jellies that were collected off the coast of NJ. Near the end of the course, the kids, and sometimes the instructors, participate in an optional talent show. The most memorable show of talent was performed, in the inaugural course, by a 10-year-old boy who did 15 minutes of non-stop impromptu stand-up comedy. Move over Jerry Seinfeld!

picture of GFP
comb jellies

THE FIELD TRIP:  The scientific component of the course is not limited to work with GFP. On July 15, for both the morning and afternoon sections, there is an all-day motor coach trip (8:00 am to 5:30 pm) to South Jersey (Stone Harbor), where naturalists will take everyone on a flat-bottom boat cruise (on the Skimmer) through the salt marsh back bay. We will pull a large plankton net through the bay, hoping to collect some jellyfish or other marine creatures. Later, a naturalist will guide the group on a short walk through sand dunes leading to the ocean beach.

REGISTRATION:  Eligibility for the course requires the submission of an essay from a list of two topics. This requirement has changed since 2017. Only one essay is required, but a bit more emphasis is being placed on the quality of the essay. As part of their eligibility, the younger students will be asked to come to campus with a parent for an interview.

THE ESSAY QUESTIONS: (answer one of the following questions to qualify for this course)

  1. The ultimate source of light energy in bioluminescent coelenterates is a molecule called coelenterazine—a type of luciferin. Oxidation of coelenterazine, under the right conditions, produces blue light (with a bioluminescence quantum yield of about 30%--three photons of light for every ten oxidized coelenterazine molecules. The other 70% is lost as heat. In the presence of GFP, however, the would-be blue light becomes green. But, now the quantum yield has risen to 80%--eight photons per ten oxidized coelenterazine molecules, with only 20% lost as heat. Here is the question: How is it possible for one coelenterazine molecule to make almost 3-times as much light with GFP present than without GFP? The laws of physics tell us that energy cannot be created, yet, in this case, it seems to be.

  2. Perhaps you have seen the movie, “Finding Nemo.” If so, you may remember that Nemo, the clown fish, has bright orange patches of color against a lighter background. You may also remember that the sea anemone, in which Nemo lives, is sometimes white with pink or green tentacle tips. The pink color comes from a non-fluorescent member of the GFP superfamily. Assuming that everything in nature has a function, driven by evolution, why do you think this sea anemone has evolved in this way, with its tentacle tips quite dramatically colored?

APPLICATION INSTRUCTIONS:

As soon as the essay is approved, you will be ready to begin registration. Procedural questions may be addressed to Dr. Ward at william.ward@rutgers.edu.