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GloFish are the first transgenic animals available to the American public. But what's
the biotechnology behind them?
"Seeing is believing with GloFish. They are
absolutely stunning!" The preceding is some of the
marketing material you'd read if you visited the
GloFish website (GloFish, 2008). Beauty may be in
the eye of the beholder, but nearly everyone would
agree that these firstand, so far,
onlytransgenic animals made available to the
general public in the United States (except in
California, pending a formal review of their
potential effect on the environment) are a worthy
conversation piece. A transgenic, or genetically
modified, organism is one that has been altered
Figure 1: The multicolored GloFish.
through recombinant DNA technology, which
http://www.glofish.com/
involves either the combining of DNA from
different genomes or the insertion of foreign DNA
into a genome. GloFish (Figure 1) are a type of transgenic zebrafish (Danio rerio) that have been
modified through the insertion of a green fluorescent protein (gfp) gene. Not all GloFish are green,
however. Rather, there are several gfp gene constructs, each encoding a different colored phenotype,
from fluorescent yellow to fluorescent red.
Currently, GloFish are the only recombinant-DNA animal that has been approved for human "use" by
the U.S. Food and Drug Administration. Their approval has raised important questions about whether,
and to what extent, genetically modified animals should be made available to consumers. But how
were scientists able to create these engineered organisms in the first place? Like so many genetic
technologies used today, recombinant DNA technology had its origins in the late 1960s and early
1970s. By the 1960s, scientists had already learned that cells repair DNA breaks by reuniting, or
recombining, the broken pieces. Thus, it was only a matter of time before researchers identified the
raw biological ingredients necessary for recombination, figured out how those ingredients functioned
together, and then tried to govern the recombining process themselves.
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Prior to these classic experiments, the idea that the genetic material was a specific chemical that
could be modified and transferred into cells was certainly controversial. But before the explosion in
recombinant DNA could begin, scientists would have to learn not only how to transfer DNA, but also
how to isolate and modify individual genes.
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cells decades earlier, this "transformation" was very inefficient, and it involved "natural" rather than
manipulated DNA. Only in the 1970s did scientists begin to use vectors to efficiently transfer genes
into bacterial cells. The first such vectors were plasmids, or small DNA molecules that live naturally
inside bacterial cells and replicate separately from a bacterium's chromosomal DNA.
Plasmids' utility as a DNA shuttle, or vector, was discovered by Stanford University biochemist Stanley
Cohen. Scientists had already established that some bacteria had what were known as antibiotic
resistance factors, or R factor-plasmids that replicated independently inside the bacterial cell. But
scientists knew little about how the different R factor genes functioned. Cohen thought that if there
were an experimental system for transforming host bacterial cells with these R-factor DNA molecules,
he and other researchers might be able to better understand R-factor biology and figure out exactly
what it was about these plasmids that made bacteria antibiotic-resistant. He and his colleagues
developed that system by demonstrating that calcium chloride-treated E. coli can be genetically
transformed into antibiotic-resistant cells by the addition of purified plasmid DNA (in this case,
purified R-factor DNA) to the bacteria during transformation (Cohen et al., 1972).
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sugar metabolism.) The significance of their achievement was its demonstration that recombinant
DNA technologies could be applied to essentially any DNA sequences, no matter how distantly related
their species of origin. In their words, these researchers "developed biochemical techniques that are
generally applicable for joining covalently any two DNA molecules" (Jackson et al., 1972). While the
scientists didn't actually introduce foreign DNA into a mammalian cell in this experiment, they
provided (proved) the means to do so.
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Jackson, D. A., et al. Biochemical method for inserting new genetic information into DNA of simian
virus 40: Circular SV40 DNA molecules containing lambda phage genes and the galactose operon of
Escherichia coli. Proceedings of the National Academy of Sciences 69, 29042909 (1972)
Kiermer, V. The dawn of recombinant DNA. Nature Milestones: DNA Technologies,
http://www.nature.com/milestones/miledna/full/miledna02.html (2007) (link to article)
Miller, H. I. FDA on transgenic animalsA dog's breakfast? Nature Biotechnology 26, 159160 (2008)
(link to article)
Zimmerman, S. B., et al. Enzymatic joining of DNA strands: A novel reaction of diphosphopyridine
nucleotide. Proceedings of the National Academy of Sciences 57, 18411848 (1967)
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