BROCK BIOL. OF MICROORGAN.-W/ACCESS >I
BROCK BIOL. OF MICROORGAN.-W/ACCESS >I
15th Edition
ISBN: 9780134810058
Author: MADIGAN
Publisher: PEARSON
bartleby

Videos

Textbook Question
Chapter 12, Problem 1AQ

Suppose you have just determined the DNA base sequence for an especially strong promoter In Escherichia coli and you are Interested in incorporating this sequence into an expression vector. Describe the steps you would use. What precautions are necessary to be sure that this promoter actually works as expected in its new location?

Expert Solution & Answer
Check Mark
Summary Introduction

To discuss:

The incorporation of the sequence of a strong promoter in E. coli into an expression vector. Steps and precautions required to evaluate the expression of the promoter in its new location.

Concept introduction:

An expression vector is a plasmid or virus and it used for gene expression. In gene expression, the gene of interest is inserted into the expression vector, which is used to carry the gene to the host. The gene encoding protein is produced in the host organism.

Explanation of Solution

  • The expression vector is used for high level gene expression of cloned genes (for example, eukaryotic genes) in prokaryotes.
  • The expression vector should contain an origin of replication, marker gene, and multiple cloning sites.
  • The promoter sequence must be incorporated into the expression vector.
  • The expression vector should contain restriction site, where the gene of interest is inserted.
  • It should help to synthesize the target protein molecule by producing the stable corresponding mRNA molecules. Therefore, strong promoter is required for binding of the RNA polymerase enzyme and that may lead to the high level of transcription.
  • The promoter should control and regulate the expression of the cloned gene, because more production of foreign protein molecules may disturb the host (E. coli) and it is considered as an important precaution. In addition, this vector should contain the transcription termination region for proper mRNA production. The promoter must contain an operator sequence in its upstream region and that provide RNA polymerase binding on the promoter region.

Want to see more full solutions like this?

Subscribe now to access step-by-step solutions to millions of textbook problems written by subject matter experts!
Students have asked these similar questions
If a soluble protein cannot be obtained from a bacterial expression vector, how else can you produce the desired protein?
Why are cDNA libraries desirable for the expression of eukaryotic genes in prokaryotes?
Describe the difference between a transcriptional fusion and a translational reporter gene fusion.  What sequences should you include in each?  What types of information can be garnered from their analyses?  Please include annotated pictures of the reporter gene expression patterns.

Chapter 12 Solutions

BROCK BIOL. OF MICROORGAN.-W/ACCESS >I

Ch. 12.3 - Prob. 3MQCh. 12.3 - Prob. 1CRCh. 12.4 - How can site-directed mutagenesis be useful to...Ch. 12.4 - What is used to alter more than a few base pairs...Ch. 12.4 - What are knockout mutations?Ch. 12.4 - What does site-directed mutagenesis allow you to...Ch. 12.5 - What is a reporter gene? The product of which...Ch. 12.5 - Prob. 2MQCh. 12.5 - Describe two widely used reporter genes.Ch. 12.6 - Prob. 1MQCh. 12.6 - Prob. 2MQCh. 12.6 - Prob. 3MQCh. 12.6 - Prob. 1CRCh. 12.7 - Prob. 1MQCh. 12.7 - Give an example of a genetically modified plant...Ch. 12.7 - How have transgenic salmon been engineered to...Ch. 12.7 - What is the Ti plasmid and how has it been of use...Ch. 12.8 - Explain why recombinant vaccines might be safer...Ch. 12.8 - Prob. 2MQCh. 12.8 - Prob. 3MQCh. 12.8 - What is a subunit vaccine and why are subunit...Ch. 12.9 - Explain why metagenomic cloning gives large...Ch. 12.9 - What types of environments are often sampled to...Ch. 12.9 - Prob. 3MQCh. 12.9 - How has metagenomics been used to find novel...Ch. 12.10 - How has Caldicellulosiruptor been modified to...Ch. 12.10 - Prob. 2MQCh. 12.10 - What has been the limiting factor in engineering...Ch. 12.10 - Prob. 1CRCh. 12.11 - What are biobricks?Ch. 12.11 - Prob. 2MQCh. 12.11 - How was Escherichia coli modified to produce a...Ch. 12.11 - Prob. 1CRCh. 12.12 - Prob. 1MQCh. 12.12 - Prob. 2MQCh. 12.12 - How is recombinant DNA inserted into a genome...Ch. 12.12 - How has the CRISPR editing technology been applied...Ch. 12.13 - Prob. 1MQCh. 12.13 - How can a tRNA be engineered to encode for a...Ch. 12.13 - Prob. 3MQCh. 12.13 - What are some mechanisms for controlling a...Ch. 12 - Suppose you have just determined the DNA base...Ch. 12 - Prob. 2AQCh. 12 - Prob. 3AQCh. 12 - Describe how you could recode Escherichia coli to...
Knowledge Booster
Biology
Learn more about
Need a deep-dive on the concept behind this application? Look no further. Learn more about this topic, biology and related others by exploring similar questions and additional content below.
Similar questions
  • Some vectors used in cloning experiments contain bacterial promotersthat are adjacent to unique cloning sites. This makes it possible to insert a gene sequence next to the bacterial promoter and express the gene in bacterial cells. These vectors are called expression vectors. If you wanted to express a eukaryotic protein in bacterial cells, would you clone genomic DNA or cDNA into the expression vector? Explain your choice.
    You then make a screen to identify potential mutants (shown as * in the diagram) that are able to constitutively activate Up Late operon in the absence of Red Bull and those that are not able to facilitate E. Coli growth even when fed Red Bull. You find that each class of mutations localize separately to two separate regions. For those mutations that prevent growth even when fed Red Bull are all clustered upstream of the core promoter around -50 bp. For those mutations that are able to constitutively activate the operon in the absence of Red Bull are all located between the coding region of sleep and wings. Further analysis of each DNA sequence shows that the sequence upstream of the promoter binds the protein wings and the region between the coding sequence of sleep and wings binds the protein sleep. When the DNA sequence of each is mutated, the ability to bind DNA is lost. Propose a final method of gene regulation of the Up Lateoperon using an updated drawn figure of the Up Late…
    Please indicate whether the following statement is true or false In eukaryotes, promoter escape involves dephosphorylation of RNA polymerase II enzyme
    • SEE MORE QUESTIONS
    Recommended textbooks for you
  • Human Anatomy & Physiology (11th Edition)
    Biology
    ISBN:9780134580999
    Author:Elaine N. Marieb, Katja N. Hoehn
    Publisher:PEARSON
    Biology 2e
    Biology
    ISBN:9781947172517
    Author:Matthew Douglas, Jung Choi, Mary Ann Clark
    Publisher:OpenStax
    Anatomy & Physiology
    Biology
    ISBN:9781259398629
    Author:McKinley, Michael P., O'loughlin, Valerie Dean, Bidle, Theresa Stouter
    Publisher:Mcgraw Hill Education,
  • Molecular Biology of the Cell (Sixth Edition)
    Biology
    ISBN:9780815344322
    Author:Bruce Alberts, Alexander D. Johnson, Julian Lewis, David Morgan, Martin Raff, Keith Roberts, Peter Walter
    Publisher:W. W. Norton & Company
    Laboratory Manual For Human Anatomy & Physiology
    Biology
    ISBN:9781260159363
    Author:Martin, Terry R., Prentice-craver, Cynthia
    Publisher:McGraw-Hill Publishing Co.
    Inquiry Into Life (16th Edition)
    Biology
    ISBN:9781260231700
    Author:Sylvia S. Mader, Michael Windelspecht
    Publisher:McGraw Hill Education
  • Human Anatomy & Physiology (11th Edition)
    Biology
    ISBN:9780134580999
    Author:Elaine N. Marieb, Katja N. Hoehn
    Publisher:PEARSON
    Biology 2e
    Biology
    ISBN:9781947172517
    Author:Matthew Douglas, Jung Choi, Mary Ann Clark
    Publisher:OpenStax
    Anatomy & Physiology
    Biology
    ISBN:9781259398629
    Author:McKinley, Michael P., O'loughlin, Valerie Dean, Bidle, Theresa Stouter
    Publisher:Mcgraw Hill Education,
    Molecular Biology of the Cell (Sixth Edition)
    Biology
    ISBN:9780815344322
    Author:Bruce Alberts, Alexander D. Johnson, Julian Lewis, David Morgan, Martin Raff, Keith Roberts, Peter Walter
    Publisher:W. W. Norton & Company
    Laboratory Manual For Human Anatomy & Physiology
    Biology
    ISBN:9781260159363
    Author:Martin, Terry R., Prentice-craver, Cynthia
    Publisher:McGraw-Hill Publishing Co.
    Inquiry Into Life (16th Edition)
    Biology
    ISBN:9781260231700
    Author:Sylvia S. Mader, Michael Windelspecht
    Publisher:McGraw Hill Education
    QCE Biology: Introduction to Gene Expression; Author: Atomi;https://www.youtube.com/watch?v=a7hydUtCIJk;License: Standard YouTube License, CC-BY