Question

Genetic Material of an organism can
be fragmented by which enzyme to
obtain gene of interest ?
(2)
ů Ligases
(n) Proteases
(in Nucleases
(iv) Polymerases​

Answer 1

Answer:

Enzymes used in molecular biology: a useful guide

Laure Rittié and Bernard Perbal

Additional article information

Abstract

Since molecular cloning has become routine laboratory technique, manufacturers offer countless sources of enzymes to generate and manipulate nucleic acids. Thus, selecting the appropriate enzyme for a specific task may seem difficult to the novice. This review aims at providing the readers with some cues for understanding the function and specificities of the different sources of polymerases, ligases, nucleases, phosphatases, methylases, and topoisomerases used for molecular cloning. We provide a description of the most commonly used enzymes of each group, and explain their properties and mechanism of action. By pointing out key requirements for each enzymatic activity and clarifying their limitations, we aim at guiding the reader in selecting appropriate enzymatic source and optimal experimental conditions for molecular cloning experiments.

Keywords: Enzymes, Molecular biology, Molecular cloning

Introduction

At a time when molecular cloning has become routine laboratory technique, we thought it was important to provide readers with some cues for understanding the function and specificities of the different enzymes used to generate and manipulate nucleic acids. Over the past few years, the tremendous expansion of cloning techniques and applications has triggered an enormous interest from laboratory suppliers. As a result, countless sources of enzymes are now available, and selecting the appropriate enzyme for a specific task may seem difficult to the novice.

Nucleic acids used for molecular cloning can be of natural or synthetic origin, and their length ranges from a few to several thousands nucleotides. Nucleic acids can be extensively manipulated, in order to acquire specific characteristics and properties. Such manipulations include propagation, ligation, digestion, or addition of modifying groups such as phosphate or methyl groups. These modifications are catalyzed by polymerases, ligases, nucleases, phosphatases, and methylases, respectively. In this review, we provide a description of the main enzymes of each group, and explain their properties and mechanism of action. Our goal is to give the reader a better understanding of the fundamental enzymatic activities that are used in molecular cloning

Answer 2

The genetic material of an organism can be fragmented by nuclease enzyme to obtain a gene of interest.

• DNA nucleases catalyze the cleavage of phosphodiester bonds.
• These enzymes play vital roles in various DNA repair processes, which involve DNA replication, base excision repair, nucleotide excision repair, mismatch repair, and double-strand break repair.

• This is a restriction enzyme which cuts the DNA in a particular site in sequences.

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