Chapter 8, Problem 18P

Summary Introduction

To analyze:

A $\text{3}\text{.5-kb}$ 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. $\text{3}\text{.5kb}$ fragment plus TFIIB and TFIID.

b. $\text{3}\text{.5kb}$ fragment plus TFIIB, TFIID, TFIIF, & RNA polymerase II.

c. $\text{3}\text{.5kb}$ fragment alone.

d. $\text{3}\text{.5kb}$ fragment plus RNA polymerase II.

e. $\text{3}\text{.5kb}$ 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.