EBK GENETICS: FROM GENES TO GENOMES
EBK GENETICS: FROM GENES TO GENOMES
6th Edition
ISBN: 9781260041255
Author: HARTWELL
Publisher: MCGRAW HILL BOOK COMPANY
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Chapter 16, Problem 36P
Summary Introduction

a.

To draw:

The recombinant plasmid that indicates the order of the four components and their arrangement in relation to the plasmid vector.

Introduction:

Erythropoietin, which is a human protein stimulates the production of red blood cells. The idea is to make a bacterium that produces this protein, and this recombinant bacterium can be used for the treatment of anemia patients.

Summary Introduction

b.

To determine:

The element that encodes the ribosome binding site for the mRNA making the fusion protein.

Introduction:

The recombinant plasmid was made to contain coding sequence for human erythropoietin, the regulatory sequence of the lac operon, a sequence encoding E.coli maltose-binding protein, and a sequence encoding a series of five amino acids. These sequences were transformed in the recombinant plasmid to induce the expression of a tagged fusion protein N MBP-DDDDK-erythropoietin C.

Summary Introduction

c.

To determine:

The elements out of given four that should be placed in the same reading frame with respect to each other.

Introduction:

The four elements present in the recombinant DNA molecule include coding sequence for erythropoietin, a sequence encoding the maltose-binding protein in E.coli and a series of five amino acids, and the regulatory sequence for lac operon.

Summary Introduction

d.

To determine:

Whether the erythropoietin coding sequences can be obtained from a human genomic DNA or from a human cDNA clone.

Introduction:

The creation of cDNA clones occurs in three major steps known as the synthesis, cloning, and validation. The cDNA clones contain open reading frames and untranslated regions.

Summary Introduction

e.

To determine:

The compound that could be used for inducing expression of the fusion protein and giving a reason about adding the compound to the medium before seeding with E.coli cells or after reaching the population of cells to high density.

Introduction:

The fusion protein that has to be made in the recombinant E.coli producing erythropoietin protein is N MBP-DDDDK-erythropoietin C.

Summary Introduction

f.

To determine:

The process by which fusion protein can be purified away from the other E.coli proteins.

Introduction:

The cells that express the fusion protein also contain many other E.coli proteins. It is considered to be important to purify the drugs away from the contaminants. It is known that MBP tightly binds to the sugar maltose and this maltose can be attached to an insoluble resin.

Summary Introduction

g.

To determine:

The way by which enterokinase can be used to separate the erythropoietin away from the rest of the fusion protein and then perform the purification of the desired pharmaceutical.

Introduction:

It is been studied that the human erythropoietin should not be attached to any other amino acid sequence. So, a protein known as enterokinase is used to cleave the proteins just C-terminal to DDDDK.

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Chapter 16 Solutions

EBK GENETICS: FROM GENES TO GENOMES

Ch. 16 - Mutants were isolated in which the constitutive...Ch. 16 - Suppose you have six strains of E. coli. One is...Ch. 16 - The previous problem raises some interesting...Ch. 16 - For each of the E. coli strains containing the lac...Ch. 16 - For each of the following growth conditions, what...Ch. 16 - For each of the following mutant E. coli strains,...Ch. 16 - Maltose utilization in E. coli requires the...Ch. 16 - Seven E. coli mutants were isolated. The activity...Ch. 16 - Cells containing missense mutations in the crp...Ch. 16 - Six strains of E.coli mutants 16 that had one of...Ch. 16 - a. The original constitutive operator mutations in...Ch. 16 - In an effort to determine the location of an...Ch. 16 - Prob. 23PCh. 16 - The footprinting experiment described in Fig....Ch. 16 - Why is the trp attenuation mechanism unique to...Ch. 16 - a. How many ribosomes are required at a minimum...Ch. 16 - The following is a sequence of the leader region...Ch. 16 - For each of the E. coli strains that follow,...Ch. 16 - Prob. 29PCh. 16 - For each element in the list that follows,...Ch. 16 - Among the structurally simplest riboswitches are...Ch. 16 - Great variation exists in the mechanisms by which...Ch. 16 - Many genes whose expression is turned on by DNA...Ch. 16 - In 2005, Frederick Blattner and his colleagues...Ch. 16 - The E.coli MalT protein is a positive regulator of...Ch. 16 - Prob. 36PCh. 16 - Prob. 37PCh. 16 - Prob. 38PCh. 16 - Prob. 39PCh. 16 - Prob. 40PCh. 16 - Prob. 41PCh. 16 - The researchers who investigated bioluminescence...Ch. 16 - Prob. 43PCh. 16 - Quorum sensing controls the expression of...Ch. 16 - Scientists are currently screening a chemical...
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