7. The coding region of the human insulin gene includes an (endoplasmic reticulum) ER signal sequence. a. Why is the presence of the ER signal sequence necessary? You wish to express human insulin in a bacterial cell using recombinant DNA technology. Steps of the procedure include: 1. insert the human insulin gene into a plasmid, 2. transform the plasmid into bacteria cells, 3. grow transformed bacterial cells in culture conditions that allow for expression of the insulin gene, 4. collect the cells and lyse them for purification of the desired human insulin protein. b. Regarding the first step, is it necessary to include the ER signal sequence in the DNA fragment coding for the human insulin gene? Briefly explain your answer. c. Why is cloning with restriction enzymes that leave sticky ends more efficient than cloning with restriction enzymes that leave blunt ends? d. In step 2 you need to select for cells that have taken up the plasmid DNA. How can you differentiate between cells with and without plasmid DNA?
7. The coding region of the human insulin gene includes an (endoplasmic reticulum) ER signal sequence. a. Why is the presence of the ER signal sequence necessary? You wish to express human insulin in a bacterial cell using recombinant DNA technology. Steps of the procedure include: 1. insert the human insulin gene into a plasmid, 2. transform the plasmid into bacteria cells, 3. grow transformed bacterial cells in culture conditions that allow for expression of the insulin gene, 4. collect the cells and lyse them for purification of the desired human insulin protein. b. Regarding the first step, is it necessary to include the ER signal sequence in the DNA fragment coding for the human insulin gene? Briefly explain your answer. c. Why is cloning with restriction enzymes that leave sticky ends more efficient than cloning with restriction enzymes that leave blunt ends? d. In step 2 you need to select for cells that have taken up the plasmid DNA. How can you differentiate between cells with and without plasmid DNA?
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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Bacterial Genomics
The study of the morphological, physiological, and evolutionary aspects of the bacterial genome is referred to as bacterial genomics. This subdisciplinary field aids in understanding how genes are assembled into genomes. Further, bacterial or microbial genomics has helped researchers in understanding the pathogenicity of bacteria and other microbes.
Transformation Experiment in Bacteria
In the discovery of genetic material, the experiment conducted by Frederick Griffith on Streptococcus pneumonia proved to be a stepping stone.
Plasmids and Vectors
The DNA molecule that exists in a circular shape and is smaller in size which is capable of its replication is called Plasmids. In other words, it is called extra-chromosomal plasmid DNA. Vectors are the molecule which is capable of carrying genetic material which can be transferred into another cell and further carry out replication and expression. Plasmids can act as vectors.
Question
Transcribed Image Text:7. The coding region of the human insulin gene includes an (endoplasmic reticulum) ER
signal sequence.
a. Why is the presence of the ER signal sequence necessary?
You wish to express human insulin in a bacterial cell using recombinant DNA technology.
Steps of the procedure include: 1. insert the human insulin gene into a plasmid, 2.
transform the plasmid into bacteria cells, 3. grow transformed bacterial cells in culture
conditions that allow for expression of the insulin gene, 4. collect the cells and lyse them
for purification of the desired human insulin protein.
b. Regarding the first step, is it necessary to include the ER signal sequence in the
DNA fragment coding for the human insulin gene? Briefly explain your answer.
c. Why is cloning with restriction enzymes that leave sticky ends more efficient
than cloning with restriction enzymes that leave blunt ends?
d. In step 2 you need to select for cells that have taken up the plasmid DNA. How
can you differentiate between cells with and without plasmid DNA?
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