You are studying a human gene, and try to express the protein in E. coli bacterial cells. To do this, you use lab cloning techniques to create an expression vector - a circular piece of DNA (plasmid) which can replicate within E. coli, and which contains an appropriate E. coli promoter sequence before the human protein sequence. Note that the sequence used is the final mRNA protein coding sequence, with all introns removed. You can detect that some of the protein is produced, but it's a very small amount. Luckily you included a positive control, which uses the same expression vector, and find that the control E. coli protein is expressed to high levels under the same conditions. A colleague suggests that you transform your same expression vector into a 'humanized' strain of E. coli that is optimized for the expression of human recombinant proteins (this is a real thing!). You do this and see protein expression – woohoo! If the genetic code is universal, why might a human gene be poorly translated in wild-type E. coli but could be better translated in the optimized strain? Propose a specific type of change which was made in the modified E. coli to account for the difference?

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
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You are studying a human gene, and try to express the protein in E. coli bacterial cells. To do this, you use lab cloning techniques to create an expression vector - a circular piece of DNA (plasmid) which can replicate within E. coli, and which contains an appropriate E. coli promoter sequence before the human protein sequence. Note that the sequence used is the final mRNA protein coding sequence, with all introns removed.


You can detect that some of the protein is produced, but it's a very small amount. Luckily you included a positive control, which uses the same expression vector, and find that the control E. coli protein is expressed to high levels under the same conditions. A colleague suggests that you transform your same expression vector into a 'humanized' strain of E. coli that is optimized for the expression of human recombinant proteins (this is a real thing!). You do this and see protein expression – woohoo!


If the genetic code is universal, why might a human gene be poorly translated in wild-type E. coli but could be better translated in the optimized strain? Propose a specific type of change which was made in the modified E. coli to account for the difference?

Expert Solution
Step 1: Introduction to cloning

Cloning is the process of creating a genetically identical copy of a biological entity. Cloning can be used to copy cells, tissues, or even entire organisms.

There are two main types of cloning: natural cloning and artificial cloning.

Natural cloning occurs naturally in many organisms, including bacteria, plants, and some animals. For example, bacteria reproduce by binary fission, which is a type of natural cloning. Plants can also reproduce by cloning through vegetative propagation, which is the process of producing new plants from existing plants without the need for sexual reproduction.

Artificial cloning is the process of creating a cloned organism using scientific techniques. There are a number of different artificial cloning techniques, including somatic cell nuclear transfer (SCNT) and molecular cloning.

Molecular cloning is the process of cloning DNA fragments.

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