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Genetic Models of Disease Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104
Submitted March 27, 2008; Revised December 2, 2008; Accepted February 16, 2009
Monitoring Editor: Nancy Moreno
| ABSTRACT |
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| INTRODUCTION |
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The OSP was originally based on the author's intuitive sense as a professional scientist that teachers should be comfortable with both the conceptual and technical content of key scientific discoveries and with the cognitive practices that scientists used when they made those discoveries (Silverman, 2003). This was essentially an historical perspective meant to engage science teachers in authentic (re)discovery. The author, as mentor, would provide technical guidance with explicit attention to the cognitive context of experiments when they were first done.
This last point is worth elaboration. Schwartz et al. (2004) have convincingly argued that for teachers and students to understand the nature of science, broadly defined, inquiry alone is insufficient, i.e., doing experiments does not necessarily illuminate the underlying epistemology. Rather, Schwartz et al. (2004) concluded that the cognitively important features of science included under the broad topic "nature of science" must be explicitly addressed, preferably in concert with inquiry, but in any case explicitly.
What are these cognitively important features? To find out, Schwartz and Lederman (2007) surveyed 24 professional scientists from several disciplines about their views of the nature of science. Notwithstanding considerable differences in detail, there was broad agreement that scientific knowledge and the process by which that knowledge is acquired are "tentative," "empirical," "theory-laden," and "creative" (Schwartz and Lederman, 2007). All true, to be sure, but such descriptors are not particularly useful formative guides to how an effective professional development program might operate.
| SPECIFIED IGNORANCE AND PEER INTERACTIONS |
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Interestingly, specified ignorance is often mentioned by scientists and educators. Richard Feynman, the physicist and renowned teacher, referred to "... the great value of a satisfactory philosophy of ignorance" (Feynman, 1998, p. 28). The biologist Joshua Lederberg suggested "... ignorance ... rather than knowledge as the terrain for learning and discovery" (Witte, 2008). James Seago, discussing the role of research in undergraduate institutions, also referred to specified ignorance using the phrase "the frontier of knowledge" (Seago, 1992).
In practice, specified ignorance almost always entails ongoing peer interactions to decide where the frontier of knowledge lies and what must be done to push it back. In the OSP, peer interactions were virtually guaranteed by the practice of having teachers work together in the same laboratory on similar research projects. The result was almost continuous discussion not only among teachers but also between teachers and project staff. Invariably, these reflective discussions included issues related to students and classrooms.
At its present state of evolution, the research dynamic is composed of three operational elements: specified ignorance, peer interactions, and gateway experiments (which will be discussed later). Importantly, specified ignorance and peer interactions are roughly equivalent to the broadly acknowledged features of the "nature of science" revealed in the survey by Schwartz and Lederman (2007). Thus, specified ignorance through peer interactions presupposes that science is empirical; that experimental data are tentative to one degree or another; that in consequence they usually need to be interpreted; that interpretations are theory-laden and consequently negotiated; and that negotiated meaning is a prerequisite for specified ignorance and hence for determining what the next experiments should be (Schwartz et al., 2004). This equivalence validates the use of Merton's three operational practices in an educational context.
The research dynamic depends critically on a scientist/mentor, whose principal pedagogical task is to guide, but not direct, teachers' research activities and to provide intellectual context. The essential feature of teacher–mentor interactions is reciprocity. Each party must be acknowledged as an expert, albeit in different subjects directly related to science education. The scientist/mentor uses his or her professional experience to convey to teachers the essential features of the research dynamic while accommodating each teacher's interests, abilities, and learning preferences, just as the teachers must accommodate diversity in their classrooms. The teachers then have the opportunity to shape the research dynamic according to their requirements as educators. This reciprocity differentiates interactions between scientist and teacher from those between scientist and graduate student: the teachers are already professionals.
| GATEWAY EXPERIMENTS |
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The historical aspects of a gateway approach are finding their way into the mainstream of educational theory (Egan, 1997; Donovan and Bransford, 2005, pp. 573–574), notwithstanding a deep resistance to using such an approach as a pedagogical tool in precollege science instruction (e.g., Duschl, 1989). Our experience has been that teachers respond well to the gateway approach (McCarty, 2003), largely because it is based on authentic, historically grounded experiments, as opposed to sterile textbooks and contrived laboratory exercises. We show below that the approach generally increases teachers' confidence in their preparedness to teach science effectively. Others have successfully tied an historical approach to contemporaneous analytical methods (Pedulla et al., 2003; Hanauer et al., 2006), although this may not always be possible.
Carried over to the classroom, gateway experiments could have pedagogical value as a lever to reconfigure the central concepts around which students organize their knowledge. We all tend to organize information in a manner that fits comfortably with our assumptions about the world. When those assumptions later prove to be wrong, as they often do, the result is confusion about and distrust of how knowledge, including scientific knowledge, is acquired and used (Bloom and Weisberg, 2007). It is well established that students bring to the classroom a remarkable body of misinformation that is notoriously difficult to change (Posner et al., 1982; Clough and Wood-Robinson, 1985; Carey, 1986; Kindfield, 1991; Pine et al., 2001; Bloom and Weisberg, 2007). In a recent interview, the physicist Lawrence Krauss (www.seedmagazine.com/news/2007/12/natalie_jeremijenko_lawrence_k.php) defined the core scientific experience this way: "... to have some cherished notion that you absolutely believe to be true proved false." Students might accomplish this through gateway experiments while guided by confident and knowledgeable teachers.
| SUMMER RESEARCH INSTITUTES |
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The following example illustrates the research dynamic as it was used in one SRI. The experiments shown were not demonstrations or in any way contrived. The teachers did these experiments themselves, motivated by peer interactions (including the author as scientist/mentor) and learning necessary techniques as they were needed. The interpersonal dynamics of the interactions are difficult to convey in the space available for this essay. Verbatim exchanges that illustrate such dynamics still require context and interpretation to mean anything. Dr. Robbie McCarty has already published such exchanges from a different SRI along with detailed contextual information, analysis, and interpretation (McCarty, 2003). Here, italicized passages are used to emphasize the author's role in providing context according to Merton's three practices. These passages were not components of lectures or in any other respect one-sided. Rather, they were the author's contributions to sometimes extended discussions in which the teachers were full participants (McCarty, 2003). Note especially how the comments followed the history of the topic, how they always entailed an experiment, and how the experiments built up a coherent body of knowledge.
Establishing the Phenomenon
The chief SRI content focus was on genes, mutations, and heredity. Our scientific understanding of these phenomena has informed a coherent view of life's history on this planet that ranks among humanity's fundamental intellectual achievements (Dobzhansky, 1973; Ayala, 2008). Yet, despite their central importance, these phenomena are among the least well understood and poorly taught biology topics (Haury, 1996, and references therein).
Genetic variation within populations, arising chiefly from a combination of mutation and recombination, is the driving force of evolution and an indispensable concept for understanding it. As a phenomenon, it is easy to see variability in evident attributes just by looking at a room full of people, but there is no simple way to find out how much, if any, of this variability is genetic (i.e. heritable). Humans are clearly not the optimal experimental system to study this question. Mendel famously used pea plants to study stable genetic variants (mutants), but, as a monk, he had few distractions and enjoyed the luxury of time.
Choosing the Experimental System
What is called for is an organism that is easy and inexpensive to grow and maintain; one that replicates quickly; and, most important because we are interested in heritability, one where the progeny of individuals (or pairs of individuals) can be easily tracked. The history of biochemical genetics suggested that microorganisms, in particular bacteria or fungi, would be the optimal choice.
Wild-type strains of the harmless bacterium Escherichia coli are able to grow rapidly on a fully defined medium consisting of salts and glucose. These cells manufacture from these few components the thousands of compounds essential for their growth and replication. As each cell grows and divides, it forms a colony of cells just like itself. Each colony, containing
108 progeny cells after 24–48 h, is a clone derived from a single founder bacterium (Figure 1).
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Specifying Ignorance
With these facts and observations in hand, the phenomenon of genetic variation could be redefined more precisely.
Suppose that the biosynthesis of each essential nutrient is genetically determined. Suppose further that different genes control the synthesis of different nutrients. It follows that if one such gene is altered so that the corresponding function is lost, mutant bacteria would lose the ability to synthesize one and only one nutrient. This defect would not prevent growth on the rich medium, where all nutrients are provided exogenously, but it would on the glucose-salts minimal medium. Additionally, if there is genetic variation within a bacterial population, different cells will have lost the ability to synthesize different nutrients. Figure 1 indicates that most cells that can grow on the rich medium also can grow on the minimal medium, so any nutritional variants are likely to be rare. What we needed to do was figure out a general method to identify nutritional variants, if they exist, as cells able to form colonies on Campbell's soup but not on glucose-salts minimal medium plates.
One approach would be to test colonies arising on Campbell's soup plates for growth on minimal plates. This could easily be done with toothpicks, as the teachers had learned when they tested isolates of bacteria for antibiotic resistance earlier in the SRI.
However, whereas 10–100% of natural isolates proved to be antibiotic resistant, the frequency of nutritional variants might be much lower, making a colony-by-colony method problematic.
The author then suggested replica plating (Lederberg and Lederberg, 1952) as more suitable. UV light-irradiated cells grown overnight in Campbell's soup medium were used to prepare master plates containing
100 colonies. By replica plating, all of these could be transferred in one step onto minimal medium plates (Figure 2), thereby increasing throughput by 2 orders of magnitude. Furthermore, the frequency of mutations can be increased by induced mutagenesis, as with UV light irradiation.
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The next level of specified ignorance was easily stated in the form of a prediction:
Each of the variants identified by replica plating should have lost the ability to synthesize one and only one nutrient supplied by Campbell's soup, and this nutrient should be different for different variants. Although identifying a specific nutritional requirement seemed akin to finding a needle in a haystack, given the metabolic complexity of a cell, actually it is not. Many biosynthetic genes in bacteria are involved in the synthesis of the 20 naturally occurring amino acids. It therefore makes sense first to screen each mutant for its ability to grow in minimal medium supplemented with each of the 20 amino acids.
Each well of a 24-well microtiter plate contained minimal medium and one amino acid. Control wells contained no addition, yeast extract, an acid hydrolysate of casein, or an enzymatic digest of casein. Each well was then inoculated with a few mutant cells and the plate was incubated overnight (Figure 3).
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This mutation was designated trp-1. Another mutant, designated met-1, required methionine. A third, vit-1, did not require any amino acid but instead had lost the ability to synthesize an unspecified B vitamin (data not shown).
One of the teachers taught chemistry and chose to pursue a question related more to chemistry than to biology: Would the amount of exogenous tryptophan limit the growth of trp-1 mutant cells and, if so, might the growth yield be used to measure the amount of tryptophan in an unknown sample? This teacher began with minimal medium supplemented with 2 mg/ml tryptophan and with serial 10-fold dilutions all the way down to 20 ng/ml tryptophan. Each medium was inoculated with a few mutant cells and the cultures were allowed to grow overnight. The teachers then measured the number of viable cells per milliliter as a function of tryptophan concentration (Figure 4).
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To test whether the growth yield of bacteria might be used to determine the concentration of exogenous tryptophan, the teacher used a commercial preparation of yeast extract from Difco (Detroit, MI). (Crushed and water-extracted yeast pills from a supermarket or health food store also should work.) The experiment was otherwise identical to that with pure tryptophan (Figure 4).
With yeast extract the number of viable cells diminished even at the first dilution, indicating that tryptophan was already limiting. As the yeast extract was diluted further, the teacher observed a good linear relationship between the amount of yeast extract and the yield of cells. Furthermore, the slope of the curve was identical to that with pure tryptophan, indicating that the efficiency of tryptophan utilization was the same as that with pure tryptophan. The teacher could therefore determine the tryptophan content of yeast by how far to the right the curve was shifted relative to pure tryptophan (Figure 4, horizontal blue arrow).
The data indicated that the yeast extract contained
2% tryptophan by mass. We obtained an analysis of an unspecified lot of yeast extract from Difco. Their value for tryptophan was 1.4%, within experimental error and uncertainty about the lot number Difco analyzed.
Authenticity
An important OSP goal was that experiments should be authentic. At the outset, "authentic" had an historical slant referring largely to the experiments themselves. It described OSP experiments chosen because of the important roles they played in the development of a scientific discipline. Thus, experiments very like those of Figures 2 and 3 were foundational to the field of biochemical genetics and won George Beadle and Ed Tatum the 1958 Nobel Prize in Physiology or Medicine. Joshua Lederberg, who invented replica plating, won along with Tatum and Beadle, albeit for experiments not discussed here. And bioassays like that shown in Figure 4 were used to identify and isolate essential bacterial growth factors (Snell et al., 1939). In this sense, "authentic" could be opposed to "contrived."
In time, authentic took on an additional, pedagogical meaning, referring to the OSP experience as a whole, independent of specific content and conforming in so far as possible to the research dynamic. Thus, other SRIs have addressed viruses (Silverman, 2003), gene regulation, mutagenesis, and antibiotic resistance. All of them incorporated simple, transparent gateway experiments coupled to specified ignorance through peer interactions.
| EVALUATION |
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"[t]he main strength of the OSP lies in the model of learning that it presents. Experiential learning in a state of relaxed alertness is indeed the optimal environment for individuals to acquire knowledge at the deepest level of being. Discovering the biological principles involved in understanding genes, mutation and heredity in the same sequence as the original researchers is not a manipulation; the data collected and the questions that come to mind as one engages in these explorations flow as naturally today as they did then. For this reason utilizing experiments and techniques that literally laid the foundation for the new science of molecular biology, and therefore have historical significance, is outstanding; the fact that these same experiences provide the stuff for building intellectual bridges between the past and the cutting-edge science of today is genius."
"Another strength of the OSP lies in the project director's responsiveness to the teachers. Silverman clearly desires to assist these teachers in transforming themselves, and their own classrooms, rather than prescribing what teachers should do. It is this attitude that makes the partnership full, and goes a long way to assure the success of the program."
McCarty also considered three weaknesses:
"... fuller advantage could be taken of the cadre of mentor teachers that the OSP has established... . The program needs to continue to collect teacher-reported data, and certainly [to] construct individual case studies for each teacher. Although there is some teacher-reported information about student learning in the classroom, and certainly documented cases of students having success with science fair projects, there [are] no hard data to document an increase in student conceptual knowledge and/or attitudes about the nature of scientific endeavor."
| SUMMARY AND FUTURE DEVELOPMENTS |
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The need for further elaboration, application, and evaluation of the research dynamic does not reduce its present value to precollege science teachers. SRIs can still be a transformative experience, changing the way science teachers teach, irrespective of what specific approaches they use in their classrooms. This alone is a signal achievement.
| ACKNOWLEDGMENTS |
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| FOOTNOTES |
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| REFERENCES |
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Witte, M. (2008). Joshua Lederberg's interest in ignorance. Science 320, 1159.
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