A 3 .5-kb segment of DNA consisting complete sequence of a mouse gene is available. The DNA segment has the promoter sequence that extends beyond the polyadenylation site of the gene. The following gel bands are observed by studying the DNA with band shift assay. These conditions are to be matched with a specific gel lane of the gel. a. 3 .5kb fragment plus TFIIB and TFIID. b. 3 .5kb fragment plus TFIIB, TFIID, TFIIF, & RNA polymerase II. c. 3 .5kb fragment alone. d. 3 .5kb fragment plus RNA polymerase II. e. 3 .5kb fragment plus TFIIB. Introduction: Band shift assay is a technique to study the interactions between protein and DNA. The assay is based on non-denaturing gel electrophoresis that runs complexes of proteins and DNA through it. The migration of either DNA or RNA is compared with migration of complex of particular protein with DNA or RNA. This study helps to reveal the degree and nature of binding between them. It is also termed as electrophoretic mobility shift assay. The Band shift assay is the separation technique for proteins, DNA, and RNA. The mixture of interest is loaded onto agarose gel and observed for speed, size, and shape.

Question

Want to see the full answer?

Answer 1

Question

Chapter 8, Problem 18P

Summary Introduction

To analyze:

A 3.5-kb segment of DNA consisting complete sequence of a mouse gene is available. The DNA segment has the promoter sequence that extends beyond the polyadenylation site of the gene. The following gel bands are observed by studying the DNA with band shift assay.

These conditions are to be matched with a specific gel lane of the gel.

a. 3.5kb fragment plus TFIIB and TFIID.

b. 3.5kb fragment plus TFIIB, TFIID, TFIIF, & RNA polymerase II.

c. 3.5kb fragment alone.

d. 3.5kb fragment plus RNA polymerase II.

e. 3.5kb fragment plus TFIIB.

Introduction:

Band shift assay is a technique to study the interactions between protein and DNA. The assay is based on non-denaturing gel electrophoresis that runs complexes of proteins and DNA through it. The migration of either DNA or RNA is compared with migration of complex of particular protein with DNA or RNA. This study helps to reveal the degree and nature of binding between them. It is also termed as electrophoretic mobility shift assay. The Band shift assay is the separation technique for proteins, DNA, and RNA. The mixture of interest is loaded onto agarose gel and observed for speed, size, and shape.

Expert Solution & Answer

Want to see the full answer?

Check out a sample textbook solution

See solution

Students have asked these similar questions

Below is an EMSA showing four different reactions, A-D. In each tube there is some combination of labelled DNA probe, Protein X (the protein you are studying), and an antibody for Protein X. Identify which combination of components are found in each of the four reactions and explain how you determined that based on the molecular interactions being studied and your knowledge of gel electrophoresis. It is possible that multiple lanes have the same component(s). A B C D EMSA

A purified recombinant protein is analyzed for molecular weight by SDS-PAGE at pH 8.5. From the protein sequence deduced from the gene that was expressed in bacteria, the protein is expected to have a molecular weight of 44,000. However, the molecular weight of the protein is found by SDS-PAGE to be 52,000. Explain the reason or reasons for this difference in molecular weight. What calculation could you make to help explain this discrepancy?

Mass spectrometry is a powerful tool in proteomics. What are the four key features of a mass spectrometer? Describe briefly how MALDI and two-dimensional polyacrylamide gel electrophoresis could be used to identify a protein expressed in cancer cells but not in normal healthy cells.