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RESEARCH INTERESTS
DNA rearrangment; SV40 transformation; molecular phylogeny
Control of DNA synthesis in SV40-transformed mouse cells
Identification of insect larvae by DNA sequencing
Teaching up-to-date science in Biology lab courses
PUBLICATIONS
I have two projects associated with DNA. The first is
an investigation of spontaneous DNA rearrangements that
occur in simian virus 40 (SV40) transformed mouse cell
lines. Rearrangements occur as short tandem
duplications or deletions within or near the integrated
SV40 DNA in mouse chromosomes. The rate of
rearrangement is very high, greater than 10-3
events/cell/division, 1000 to 10,000 times higher than
the usual rate of spontaneous mutation. The high rate
requires the presence of the SV40 early control region, a
350 nucleotide segment of SV40 DNA containing the
SV40 origin of DNA replication, one or two enhancers,
and a promoter of SV40 T antigen transcription. In one
system, rearrangements required the SV40 control region
but not the T antigen gene or its expression. In another
system the rate of rearrangement was tightly correlated
with the type of T antigen expressed. Figure 1 shows
that the line X1 which expressed wild-type T antigen
rearranged at a higher rate than did the closely related
SVT2 which expressed a mutant larger T antigen. We
propose that the control region is required for
rearrangement but T antigen expression can modulate the
rate of rearrangement.
The second project concerns the molecular systematics
of insects, identification of insects by their DNA
sequences. The medical interest in this project is to
identify disease-bearing species, fleas in particular.
There are very few flea sequences in the Gen-EMBL DNA
databases so we are helping to remedy that deficiency.
To establish correspondences between morphology and
DNA sequences we collaborate with Dr. Robert E. Elbel, a
veteran entomologist. We developed a method for
extracting DNA from an insect without destroying the
exoskeleton so that morphology and sequence of a single
insect can both be studied. We use PCR to amplify
nuclear (ribosomal ITS) and mitochondrial (COI)
sequences. Sequences are variable enough to identify
species and subgroups within species. The methods are
simple and reliable enough to be used for teaching PCR
and DNA sequencing in undergraduate lab courses. In
Figure 2 a hypervariable region of sequence
discriminated between individual insects.
Selected Publications
Gurney, T. Jr., F. Hutchinson, and R.D. Young (1965) Condensation of tungsten on tungsten in atomic detail: Observation with the field-ion microscope. J. Chem. Phys. 42:3939-3942.
Gurney, T. Jr. and M.S. Fox (1968) Physical and genetic hybrids formed in bacterial transformation. J. Mol. Biol. 32:83-100.
Foster, D.N. and T. Gurney, Jr. (1976) Nuclear location of mammalian DNA polymerase activities. J. Biol. Chem. 25l:7893-7898.
Gurney, T. Jr. and M.W. Collard (1984) Nonaqueous fractionation of HeLa cells in glycols. Anal. Biochem. 139:25-34.
Gurney, T., Jr., and E.G. Gurney (1989) Spontaneous rearrangement of inte grated simian virus 40 DNA in nine transformed rodent cell lines. J. Virol. 63:165-174.
Gurney, T, Elbel, R, Ratnapradpa, D, and Bossard, R (2000) Introduction to the molecular phylogeny of insects . Pages 63-79 in Tested studies for laboratory teaching (SJ Karcher, ed), Proceedings of the 21st conference of the Association for Biology Laboratory Education (Lincoln, NE, June 5-10, 1999), 509 p.
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