Question

You are studying the part of the brain that mediates stress. You have treated some mice under stressed conditions and you also have control mice. You isolate the appropriate brain regions from both groups.

Describe what you would do to identify changes in genes associated with acetylated lysine 9 on histone H3.

What would you do to prepare the brain samples for analysis?

Would technique would you use to purify genes associated with aceylated lysine 9 histone H3 and how does it work?

How would you identify all the genes in the purified samples?

What kind of data would you get in the end that would tell you what genes changed in their association with acetylated lysine 9 histone H3 in response to stress?

ANSWER ALL please

Answer 1

1. Describe what you would do to identify changes in genes associated with acetylated lysine 9 on histone H3?

The changes in the expression levels of proteins associated with acylated lysine 9 H3 can be done by protein assay in western blotting. After collecting the brain cells, extract nuclear proteins from the cells using any commercially available protein isolation kit. Then quantify the proteins using western blot analysis by probing the proteins with corresponding antibodies.

2. What would you do to prepare the brain samples for analysis?

Remove the brain of appropriate aged mice. Dissect carefully and collect the region of the brain without any damages. immerse the sample immediately in ice-cold CSF solution to keep the cells viable. Then changed it into an appropriate buffer with suitable pH for preparing cell lysate and further protein purification.

3. Which technique would you use to purify genes associated with acylated lysine 9 histone H3 and how does it work?

The proteins can be purified by affinity chromatography in which the sample passing through a column of antibodies embedded in it. Then collect the proteins from the column and run in western blot which will resolve each protein on the basis of their mass (confirmatory step).

4. How would you identify all the genes in the purified samples?

western blot may give the mass of each protein which can be compared with the known value of possible ones.

5. What kind of data would you get at the end that would tell you what genes changed in their association with acetylated lysine 9 histone H3 in response to stress?

The final data will tell about the level of expression (will be inferred from the thickness of the band in the blot) of each protein associated with acetylated lysine 9 H3 in both experimental stressed one and the control one. so it will be easy for us to compare the protein expression of control and experimental mice in case of stree response