If the binding activity was purified, suggest two experimental approaches that could be taken to verify that this factor is in fact a transcription factor? Explain.

Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
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If the binding activity was purified, suggest two experimental approaches that could be taken to verify that this factor is in fact a transcription factor? Explain.
An electrophoretic mobility shift assay (EMSA) was performed using a radiolabeled
DNA fragment from the sequence upstream of gene X. This DNA probe was incubated
with (+) or without (-) nuclear extract isolated from tissues A (bone); B (lung); C (brain);
and D (skin). The DNA-protein complexes were then fractionated on nondenaturing
polyacrylamide gels. The gels were exposed to autoradiographic film; the results are
presented in the figure.
Nuclear -
extract
A
B
+
C
+
D
+
Transcribed Image Text:An electrophoretic mobility shift assay (EMSA) was performed using a radiolabeled DNA fragment from the sequence upstream of gene X. This DNA probe was incubated with (+) or without (-) nuclear extract isolated from tissues A (bone); B (lung); C (brain); and D (skin). The DNA-protein complexes were then fractionated on nondenaturing polyacrylamide gels. The gels were exposed to autoradiographic film; the results are presented in the figure. Nuclear - extract A B + C + D +
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