Chapter 22, Problem 1P

Interpretation Introduction

Interpretation:

The approach to determine the processivity of deoxyribonucleic acid (DNA) polymerase enzyme should be experimentally explained.

Concept Introduction:

Deoxyribonucleic acid (DNA) polymerase is an enzyme which is mainly responsible for the formation of deoxyribonucleic acid (DNA) molecule stretch from dNTPs (deoxyribonucleotides). The enzyme is critical for DNA replication and helps in synthesizing sister DNA strands from the original deoxyribonucleic acid molecule.

DNA processivity is defined as the degree of an average number of nucleotides added before the release of the enzyme deoxyribonucleic acid (DNA) polymerase from the primer. In a binding event, the processivity ranges from few nucleotides to more than $50000$ nucleotides. The fidelity and processivity of the enzyme deoxyribonucleic acid (DNA) polymerase will be high if the frequency of incorporation of the wrong base pair is low.

Explanation of Solution

The incorporation essay is the experimental approach to determine the processivity of enzyme deoxyribonucleic acid (DNA) polymerase. Below steps are involved in the experimental approach:

1. A primer, which is known as deoxyribonucleic acid (DNA) template and a mixture of labelled deoxyribonucleotides (dNTPs) are used. The DNA which is newly synthesized is labelled along with the newly incorporated deoxyribonucleotides (dNTPs) and new DNA is separated from the strand of deoxyribonucleic acid (DNA) template.
2. The reaction of the enzyme deoxyribonucleic acid (DNA) polymerase is paused after a specific time and a library of single DNA strand is formed. The labelled DNA strand is separated and various number of deoxyribonucleotides (dNTPs) can be viewed through gel electrophoresis and autoradiography methods.
3. The above techniques separated the deoxyribonucleic acid (DNA) based on its size. Also, the amount of deoxyribonucleic acid (DNA) synthesized can be calculated by determining the amount of labelled nucleotides incorporation into deoxyribonucleic acid (DNA) chain.

Thus, processivity of the enzyme deoxyribonucleic acid (DNA) polymerase can be experimentally determined through incorporation essay.