3. (5 points) From the figure below, what can you conclude about how the protein

Question

3. (5 points) From the figure below, what can you conclude about how the proteins SVP, FLM-B, and FLM-5 interact with the DNA sequence of the SEP3 gene? It is important to note the relative protein sizes: monomers of SVP are 40 kDa and monom versions of FLM proteins are 15 kDa in size. The weight of the SEP3 probe is negligible. In your answer be sure to identify which proteins are bound to the SEP3 probe DNA for each band that appears on the figure. ers of both 175 150 100 50 25 Figure 8 EMSA using SEP3 probe (32P-labeled DNA obtained by amplifying the SEP3 gene). The probe was incubated with proteins as indicated. Numbers indicate approximate sizes in kDa. SEP3 probe 4(4 points) Describe how you would perform a co-IP assay to analyze the interactions between SVP, FLM-B, and FLM-6.Be sure to describe your expected results (based on your conclusions that you made in question 4) and draw a figure depicting your expected results

Explanation / Answer

electrophoretic mobility shift assay (EMSA) also referred as a gel shift assay,  band shift assay, or gel retardation assay, is a common affinity electrophoresis technique used to study protein DNA or protein–RNA interactions

solutions of protein and nucleic acid are combined and resulting mixtures are subjected to electrophoresis under native conditions through polyacrylamide and after electrophoresis, the distribution of species containing nucleic acid is determined here usually by autoradiography of 32P-labeled nucleic acid. In simple or general view, protein nucleic acid complexes migrate more slowly than corresponding free nucleic acid

This information is suggestive of a fact that FLM beta and delta are co-regulatory proteins for the probe of DNA whereby the DNA sequence is the target for dimer of FLM delta and teteramer of FLM beta

EMSA test is used to investigate the relationship between genetic sequence or DNA and the protein in order to investigate the extent of binding of the protein with DNA and this type of experiment is ususally conducted to understand effect of a regulatory protein on gene expression.By the diagrammatic information, it can be clearly seen that protein FLM beta binds very strongly with probe thus giving 4 bands in the membrane. Other side, FLM delta binds slightly weakly with the probe thus giving only 2 bands. However, since two bands of FLM delta are similar to those of FLM beta, it's very likely that both these subunits bind to each other during gene regulation and then modulate gene expression.

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