This special, 3-credit Rutgers Summer Session lab course has been designed for younger, profoundly gifted and talented students.  The course is also recommended for early-admitted, first-year matriculated students at Rutgers University’s New Brunswick campus.  Children from 10 to 17 years of age may be eligible, but each admitted student must have taken a high school level chemistry course, an equivalent course via home schooling, or an internet-accessible introductory chemistry course now being created.  Admission is by permission of the instructor and the director of the Rutgers Summer Session program.

The curriculum will include a major hands-on biochemistry lab component, an aquarium design project, observations of live bioluminescent jellyfish and comb jellies, and an art project involving the jellyfish or comb jellies that we collect.  There will be an all-day, professionally-narrated, salt marsh, flat-bottomed boat trip along the sheltered bays near Stone Harbor in south Jersey.  There will even be an optional talent show.  While half the group is taking the boat ride, the other half will be going on a walking field trip along the Atlantic dunes, a trip led by a professional naturalist.  Later the two groups will switch from one activity to the other.   Back in the lecture room, registrants will take one or more quizzes, submit a final lab report, and take a comprehensive final exam.  In other words, this is a real college-level course for which students will register as Rutgers students and will earn three college credits.  But, considering the age of the students, a large part of the grade will be based on participation.  

Throughout the course, twenty students in groups of 5 will be lead by (and continuously supervised by) one of our four well-trained instructors.  In the lab, students will extract, purify, and characterize the recombinant green-fluorescent protein (GFP) from genetically transformed, frozen, non-pathogenic E. coli cells.  Among other techniques, the students will perform column chromatography of several types, gel electrophoresis, absorption spectrophotometry, and high pressure liquid chromatography.  Dr. Ward will do parallel experiments from time to time and share his results with the entire class.

Science will be combined with art as students will create (and take home) matted, artistic wall hangings made from real jellyfish www.marineimpressions.net.   In addition, each group will design an aquarium system  that might be suitable for maintaining the exceedingly fragile comb jellies we will collect.  Near the end of the course, students will be afforded an opportunity to participate in a talent show.   Creating their own graphs that depict the results of lab experiments can also be considered an artistic venture.

Registration information and administrative details about this course are available on-line at http://summer.rutgers.edu/highschool.     The meeting place is the Cook Campus (SEBS) biochemistry lab/lecture suite located on the second floor of Lipman Hall, 76 Lipman Drive, New Brunswick, NJ.  The course runs from 9 am to 12 noon 5 days a week for 3 weeks.  Two sections are available—Section D1 runs from June 25 to July 16 (with a break for July 4) and Section G1 runs from July 16 to August 3.  Both classes, together, will take the all-day field trip to Stone Harbor on July 16.  The combined fee for commercial bus transportation to Stone Harbor and the services of two professional naturalists totals $100.  This fee is separate from Summer Session registration.  Enrollment is normally limited to 20 students per session, although we may be able to accommodate a few more students if there is sufficient demand. 

If you have questions about course content, contact Dr. William Ward, crebb@rci.rutgers.edu.  For registration and administrative information, contact http://summer.rutgers.edu/highschool.   For direct personal information concerning eligibility or registration matters, you may contact Ms. Liz Beasley directly at houghliz@docs.rutgers.edu.

HOW TO BEGIN

1. Read the EXPANDED COURSE DESCRIPTION, (shown below), to be sure if you that are interested in attending the course.  If so, there are several steps you need to take.  First, you need to check with Summer School about admission requirements.  These requirements include submission of scores on the PSAT test (or the equivalent) and, for those under the age of 12, a personal interview may be required.   You will need to have taken a high school chemistry course or the equivalent.  Alternatively, you may satisfy the chemistry requirement with a home-schooled course in beginning chemistry, a tutorial administered by a chemistry teacher, or an introductory chemistry course that Professor Ward will soon be posting on the Internet.  Even if you have taken a chemistry course or been tutored, check the Internet chemistry course to be sure that the material posted on this site is within your grasp.  Your having satisfied the chemistry requirement needs to be reported to Professor Ward along with the essays you will submit (see paragraph 2).
   

2. Once you begin to fulfill the above requirements, you need to write three short essays (generally one page for each).  The first essay is required of everyone.  Then you chose two other topics from a list of seven.  Near the end of this document are the topics.  The essays are to be submitted directly to Professor Ward at the email address crebb@rci.rutgers.edu.   Direct emails or attachments are fine, but if you send an attachment, be sure to place your name at the top of the first page of the attachment.  I WILL NOT BE GRADING ON THE BASIS OF RIGHT OR WRONG ANSWERS.  I AM LOOKING FOR INTEREST, ZEAL, IMAGINATION, CREATIVITY, AND EXPRESSIONS OF CURIOSITY.  Partial answers to some of the questions can be found in the literature, but, if you find helpful information on the Internet, your answers still have to reflect your own insights and imagination. 
   
   
3. After I read and approve your essays, I will send you, by return email, a SPECIAL PERMISSION NUMBER required for registration.  It is unlikely that you will be rejected unless you cannot deal effectively with the questions or if the class fills before your registration is completed.  Remember that you will not be accepted into the class until all registration matters have been satisfied.  If your essays are not approved, you will be offered a second chance following my suggestions about improving your second submission.  We will keep a waiting list if registration goes over the limit.  You will be unable to register without your special permission number.
   
   
4. Other registration steps are conducted electronically, as described on the Rutgers Summer School site http://summer.rutgers.edu/highschool.

Expanded Course Description:

Section 1 of this course runs from June 25 to July 13, from 9 am until noon each day (with a break for July 4).  The second section runs from July 13 to August 2.  Registrants in the course will conduct biochemical research on jellyfish-derived green-fluorescent protein (GFP) that has been cloned into non-pathogenic E. coli cells.  GFP is now one of the most famous proteins in the world, having been the subject of more than 41,000 research papers and having been awarded the Nobel Prize in 2008. 

On July 16, (weather permitting) registrants from both sections will be transported to Stone Harbor, NJ, where we will observe a vibrant salt marsh area.  A professional naturalist will describe the animals and marsh plants of the area.  There will be an additional walking tour of the dunes along the ocean.  From the flat-bottomed boat, we will look for transparent bioluminescent comb jellies, Mnemiopsis leidyi, that are almost always present in the summer in river mouths, inlets, and bays. We may also find the larger, cloudy-white, non-stinging, saucer-shaped medusae that appear sporadically on the Jersey coast.  They are also bioluminescent.  This will be an all-day trip that requires a payment of $100 above the cost of registration.

Once back at the Biochemistry teaching lab, we will extract recombinant GFP from transformed E. coli.  We will then proceed to purify the GFP with a variety of methods including the novel three-phase partitioning step, one or more types of column chromatography, and high pressure liquid chromatography. Comb jellies (ctenophores) will be observed live in the lab where we will observe their swimming behavior, their brilliant bioluminescence, and perhaps larvae that may appear over night. We will feed them with brine shrimp larvae and, if they survive for a couple of days, we will watch how the orange color of the brine shrimp begins to color the comb jellies. All comb jellies swim by means of fused cilia—the largest cilia in the animal kingdom. If we are very lucky, we may observe and collect the predatory (and also bioluminescent) comb jelly (Beroe), little more than a swimming stomach. This predatory comb jelly loves to eat, in one huge gulp, the more gentle Mnemiopsis. We will observe ctenophore bioluminescence and the effect of bright light on comb jelly luminescence. We will watch the swimming motion of the cilia with a microscope, and with proper lighting, we will observe iridescence from the comb rows. Comb jellies reproduce every night. By the next day, there could be tiny larvae that we can observe to see a large collection of interesting photographs of luminescent animals check out our short course web site at: http://www.rci.rutgers.edu/~crebb/protein.html.

Comb jellies are very difficult to maintain in captivity. Teams of students will be asked to design aquarium systems that might be suitable for maintaining comb jellies for long periods of time. This is not an ordinary course. “Experiments with GFP: The Art and the Science” attempts to integrate aspects of biology, marine ecology, invertebrate anatomy, microscopy, physiology, biochemistry, physics, and engineering into a seamless whole. Each student will be expected to keep notes and then to write a report at the end of the session. Library and/or Internet research is encouraged. The reports will be graded and returned.

Back in the Lipman Hall teaching lab on the Cook College Campus, science will be combined with art as participants create (and take home) matted, artistic wall hangings made from real jellyfish www.marineimpressions.net.  This course is recommended for profoundly gifted younger students and early-admitted entering college students.

Registration to this course is by Special Permission Only. Special permission requires submitting three essays to Professor Ward as directed above. After receiving the Special Permission Number, students must create a user account on Summer School site and complete the application/registration form. To return to the Summer School for registration go back to http://summer.rutgers.edu/highschool.

ESSAY QUESTIONS: ONE PAGE FOR EACH IS SUFFICIENT. BE CREATIVE!

There are two parts to this assignment.  The single question in Part 1 is required of all students.  You are to write an essay explaining why you want to take this course. Part II has 7 choices.  You pick two and write short essays about those you have chosen.  Be sure to number the questions you have selected as they are numbered below. Be as creative as you can and explain, as well, why you chose to answer the questions you picked.  Don’t be put off by long questions.  Sometimes long questions are easier to answer.

PART 1. What do you feel is artistic about the practice of biochemistry?

PART II.

2. In the late 1960’s and early 1970’s, GFP was discovered in bioluminescent coelenterates within the classes: hydrozoa (jellies that alternate generations between polyps and medusae) and anthozoa (sea pansies and sea pens), but it was not seen in scyphozoa (the true jellyfish). Coelenterate “luciferin,” that has come to be known as coelenterazine, produces blue light from its excited state following oxidation. Unlike other luciferins, whose color of light can sometimes be modulated by external factors, coelenterazine is unable to generate other colors of light. Yet most bioluminescent coelenterates make green light. The actual emitter turns out to be green-fluorescent protein (GFP). For many years, it was assumed by those working closely with bioluminescent coelenterates that GFP has one specific function—to alter the color of bioluminescence from blue to green.  This color change occurs by radiationless energy transfer from excited state oxyluciferin to the GFP. Yet, in later years, it was found that GFP is part of a superfamily of closely related pigmented proteins, some fluorescent and some just brilliantly colored. Not only are these proteins found in non-bioluminescent coelenterates (corals and sea anemones) but they appear in other phyla. What does this teach you about the objectivity of scientific inquiry?

3.  Many bioluminescent coelenterates live in coastal estuaries. Speculate on why most of these coastal coelenterates have evolved (or adopted) green fluorescent proteins to create green bioluminescence.

4.  Green-fluorescent protein, in its native, properly folded, state, is highly resistant to proteases (enzymes that digest proteins). It would seem, then, that GFP is a most unlikely protein around which to build an assay for proteases. Yet, our group is doing this right now. Explain how this might be possible.

5.  Perhaps you have seen the movie “Finding Nemo.” If so, you may remember that Nemo, the clown fish, has a home among the stinging tentacles of a nearly all-white sea anemone. These, and other non-colored sea anemones, have pink-tipped tentacles. The pinkish 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 this way, with its tentacle tips quite dramatically colored?

6.  Conscientious and ethical scientists do everything in their power to publish research papers that are error-free. Sometimes mistakes happen. I published a paper years ago that had a glaring quantitative error (not really my fault as I followed the guidance of a more senior scientist). In this paper, I incorrectly reported the fluorescent quantum yield of Renilla reniformis GFP to be 0.30. Fluorescent quantum yield of a fluorescent molecule is defined as the ratio of the number of photons of light emitted by that molecule divided by the number of photons of light absorbed by that molecule. A simple way to make this measurement is by reference to a fully accepted standard. The organic molecule, fluorescein, has an accepted fluorescence quantum yield of 0.80. How would you use fluorescein as a standard in measuring the quantum yield of a fluorescent protein?

7.  Sometimes published papers have more than one error. Sometimes they are riddled with errors. See if you can find the Biochemistry paper by Karkhanis and Cormier (1971) and tell me what you think about this paper (hint—it is really, really, really bad). Alternatively, download the paper in Biochemistry, 1974, by Morise, Shimomura, Johnson, and Winant. You may recall that Osamu Shimomura received the Nobel Prize in Chemistry for his work on GFP, yet every paper he wrote on GFP has one or more errors—some are whoppers. See if you can find out what is crucially wrong with this paper. If you are really ambitious, see if you can find out what is wrong with his other papers on GFP.

8.  Coelenterazine has been known for many years to be a cyclic tripeptide made up of the amino acids Phe-Tyr-Tyr. The chromophore of GFP is also a cyclic tripeptide formed from the amino acids Ser-Tyr-Gly. I believe that some organisms that use coelenterazine for their bioluminescence reaction make this luciferin molecule de novo in the same way that GFP is made—by autocatalytic cyclization of three amino acids. Evidence comes from unpublished feeding experiments where captured jellies grown in artificial sea water INCREASE their level of coelenterazine while being fed only brine shrimp. If the mechanism for coelenterazine biosynthesis is analogous to the mechanism for biosynthesis of the GFP chromophore, then there must be a pre-luciferin protein that gives rise to coelenterazine. How would you search for this pre-luciferin protein. HINT: start with a search for the gene. Propose an elegant way to screen for the pre-luciferin gene.

Remember to send your essay answers directly by email to crebb@rci.rutgers.edu

Sincerely,
Professor William Ward