The expected results of electrophoretic mobility shift assay (EMSA) done on the following samples: Lane 1. 600 base pair (bp) fragment containinga glucocorticoid response element (GRE) and glucocorticoid receptor. Lane 2. 600 bp segment having a GRE, glucocorticoid receptor, and glucocorticoid hormone. Lane 3. 600 bp segment with a GRE and CREB (cyclic adenosine monophosphate (cAMP) response element binding protein) protein. Lane 4. 700 bp segment having a cAMP response element (CRE) and CREB protein. Lane 5. 700 bp fragment having a CRE along with CREB protein, and hormone which causes phosphorylation of CREB protein. Lane 6. 700 bp segment with a CRE as well as glucocorticoid receptor, and glucocorticoid hormone. Introduction: Electrophoretic mobility shift assay is done to study the interaction of protein and deoxyribonucleic acid (DNA) or the interaction of protein and ribonucleic acid (RNA). This technique is used to find out whether the given protein or the mixture of proteins can bind to a specific DNA or RNA fragment.

Question

Want to see the full answer?

Answer 1

Question

Chapter 21, Problem 31EQ

Summary Introduction

To review:

The expected results of electrophoretic mobility shift assay (EMSA) done on the following samples:

Lane 1. 600 base pair (bp) fragment containinga glucocorticoid response element (GRE) and glucocorticoid receptor.

Lane 2. 600 bp segment having a GRE, glucocorticoid receptor, and glucocorticoid hormone.

Lane 3. 600 bp segment with a GRE and CREB (cyclic adenosine monophosphate (cAMP) response element binding protein) protein.

Lane 4. 700 bp segment having a cAMP response element (CRE) and CREB protein.

Lane 5. 700 bp fragment having a CRE along with CREB protein, and hormone which causes phosphorylation of CREB protein.

Lane 6. 700 bp segment with a CRE as well as glucocorticoid receptor, and glucocorticoid hormone.

Introduction:

Electrophoretic mobility shift assay is done to study the interaction of protein and deoxyribonucleic acid (DNA) or the interaction of protein and ribonucleic acid (RNA). This technique is used to find out whether the given protein or the mixture of proteins can bind to a specific DNA or RNA fragment.

Expert Solution & Answer

Want to see the full answer?

Check out a sample textbook solution

See solution

Students have asked these similar questions

ATM is a kinase that phosphorylates histone H2AX in response to double-stranded DNA breaks. Which of the following scenarios would most quickly regulate ATM activity in the cell? a) Adding silencing methyl groups to cytosines in the Atm gene b) Modifying the histone code for the Atm gene c) Increasing expression of a miRNA specific for the Atm mRNA d) Activating an E3 ubiquitin ligase specific for the ATM protein

The binding of a small effector molecule, protein-protein interactions, and covalent modifications are three common ways to modulate the activities of transcription factors. Which of these three mechanisms are used by steroid receptors and by the CREB protein?

If the lacl gene is mutated so that the repressor protein no longer binds to lactose, what will be the effect on the expression of B-galactosidase in lactose's presence and absence? Explain. If the promoter for lacl is mutated so that the expression of the repressor increases, what will be the effect on the expression of B-galactosidase in the presence and absence of lactose? Explain. D. (Extremely tricky question!) Describe the behavior of the lac operon assuming that the lacl gene has been mutated so that the repressor now binds to DNA in the presence of lactose but cannot bind to DNA in the absence of lactose.