Essentials of Genetics (9th Edition) - Standalone book
Essentials of Genetics (9th Edition) - Standalone book
9th Edition
ISBN: 9780134047799
Author: William S. Klug, Michael R. Cummings, Charlotte A. Spencer, Michael A. Palladino
Publisher: PEARSON
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Chapter 17, Problem 5PDQ
Summary Introduction

To review:

The reason for use of wide host range in RDT (recombinant DNA or deoxyribonucleic acid technology) for the transformation of recombinant DNA into a bacterial cell.

Introduction:

RDT involves gene cloning and genetic engineering process, in which manipulation of the genetic material of an organism is carried out. It is done by inserting the gene of interest in the vector, which carries it to a specific host to produce multiple new genetic combinations with the help of RDT. There are many tools that are used in the gene cloning experiment, such as restriction endonucleases, DNA ligases, foreign DNA or DNA insert, vector, and the host cell. Moreover, the host cell supports the amplification of the DNA fragment for gene expression at the phenotypic level.

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Following are four processes common to most cloning experiments:  a)    transforming bacteria b)    plating bacteria on selective medium c)     cutting DNA with restriction endonucleases d)    ligating DNA fragments Place components of this list in the order in which they would most likely occur during a cloning experiment.
What are the advantages of using organisms such as bacteria and yeast as cloning hosts? What safety considerations must be made when choosing a cloning host to make proteins for human use?
With the use of well-illustrated diagrams, reconstruct the entire cloning process by explaining different stages of the cloning process that involves the following:a. Isolation of target DNA fragments (often referred to as inserts)b. Ligation of inserts into the plasmid, creating recombinant molecules c. Transformation of recombinant plasmids into bacteria or other suitable host for propagationd. Screening/selection of hosts containing the intended recombinant plasmid. For this stage(d), discuss the importance of a second marker that can be used for screening of genomic DNA for colonies containing the pka-1 under the principle of insertional inactivation. This should be properly explained using all the attributes of the plasmid described above.
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