Nucleic Acid Aptamer Selection Against GUS (Beta-Glucuronidase) for use
as a Reporter Molecule
Maneesha Gaddikoppula, Fall 2013
GUS
(Beta-Glucuronidase) is a reporter molecule that yields a colorful product when
immersed in a solution containing a specific substrate. One of the common methods
of GUS reporter assay is through histochemical staining which requires one of a
variety of colorless substrates, such as X-Gal, resulting in a blue colorful
product. Finding an aptamer against GUS can prove to be an effective diagnostic
tool in the future.
Figure 1 below illustrates the usefulness
of an aptamer against GUS. When an aptamer is bound to GUS, GUS is no longer
able to bind to the substrate (X-Gal), and thus will not form a colorful
product. However, the aptamer can be modified such that the part of the aptamer
that is not bound to GUS can bind to various other molecules specifically, such
as mRNA or other protein. When that mRNA or protein is abundant, the aptamer
will bind to that, leaving GUS open to bind to the substrate. This allows a
colorful product to form, showing that the specific protein or mRNA is present.
Had there been no mRNA or protein present, the aptamer would never have unbound
from GUS, and a colorful product would never have formed. This can make an
aptamer against GUS a useful diagnostic tool.
Specific Aim 1: Selection of an RNA aptamer against GUS.
GUS is a reporter molecule that forms a colorful product
when bound to a substrate. An aptamer can inhibit the function of GUS, and
prevent the formation of a colorful product. This process can be used as an
important diagnostic tool towards other diseases.
Figure 1 illustrates the
role of an aptamer against GUS when a specific mRNA or
protein is present that the aptamer is
modified to bind preferentially to.
Target Order
Information:
Vendor: Ellington Lab (through Jimmy H. Golliher)
Vendor Website: http://ellingtonlab.org
Vendor Telephone Number:
Central Lab
Telephone
: 1-512-471-6445
Office Manager
: 1-512-232-3426
Cost per unit: $0
Cost per round: $0
Link to Proposal:
https://www.dropbox.com/s/z2gqez447rwgfks/Gaddikoppula%2C%20Maneesha%20-%20Proposal.pdf
Link to Progress Update 1:
https://www.dropbox.com/s/v11yqcasv1fbv28/Gaddikoppula%2C%20Maneesha%20-%20Progress%20Report%201%20Fall.pdf
Link to Progress Update 2:
https://www.dropbox.com/s/6zie6zep9ta377x/Gaddikoppula%2C%20Maneesha%20-%20Progress%20Report%202%20Fall.pdf
Link to Final Progress Report:
Resources:
Jefferson RA, Burgess SM, Hirsh D. “beta-Glucuronidase
from Escherichia coli as a gene-fusion marker.” Proc Natl Acad Sci U S A. 1986;83:8447–8451.
Jefferson RA, Kavanagh TA, Bevan MW. “GUS
fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in
higher plants.” EMBO J.
1987;13:3901–3907.
Marathe SV, McEwen JE (February 1995).
"Vectors with the gus reporter gene for identifying and quantitating
promoter regions in Saccharomyces cerevisiae". Gene 154 (1): 105–7.
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