You’re working in a research lab, and your current task is to clone the gene that codes for tyrosinase from potatoes. You grind up some potato, extracts the DNA from it and digests the DNA with two different restriction enzymes (separately, not together): EcoRI and BamHI. You then obtain the cloning vector, pUC19, and digest it with the same two enzymes. You then run a gel which is shown to the right.You mix the potato DNA (digested with the enzyme you specified in part B) with cloning vector DNA (digested with the same enzyme). You then add the mixture to E. coli cells and carry out a transformation procedure so that the cells can each update a plasmid. You then plate the cells on a plate containing antibiotic. What antibiotic would be in the plate? Why would there be antibiotic in the plate? Be specific!Unfortunately, you don’t get a single bacterial colony to grow on the plate. Not even one! You review your procedure and realize that when mixing the digested potato DNA and digested plasmid that you forget to add a critical enzyme. What enzyme needed to be added? Why? Be very specific in your explanation. OOO D O||IIII | | || 2500lacza396-454Polylinker500аmpPUC192686 bp2000ori10001500

Answered: You’re working in a research lab, and…

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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Question

You’re working in a research lab, and your current task is to clone the gene that codes for tyrosinase from potatoes. You grind up some potato, extracts the DNA from it and digests the DNA with two different restriction enzymes (separately, not together): EcoRI and BamHI. You then obtain the cloning vector, pUC19, and digest it with the same two enzymes. You then run a gel which is shown to the right.

You mix the potato DNA (digested with the enzyme you specified in part B) with cloning vector DNA (digested with the same enzyme). You then add the mixture to E. coli cells and carry out a transformation procedure so that the cells can each update a plasmid. You then plate the cells on a plate containing antibiotic. What antibiotic would be in the plate? Why would there be antibiotic in the plate? Be specific!
Unfortunately, you don’t get a single bacterial colony to grow on the plate. Not even one! You review your procedure and realize that when mixing the digested potato DNA and digested plasmid that you forget to add a critical enzyme. What enzyme needed to be added? Why? Be very specific in your explanation.

2500
lacza
396-454
Polylinker
500
аmp
PUC19
2686 bp
2000
ori
1000
1500

Expert Solution

Introduction

Recombinant DNA technology is one of most significant techniques of genetic engineering. It enables us to introduce traits/characters of our desire into an organism by inserting a gene that codes for this trait into the organism. These traits or characters could be the something that enable a plant to survive in extreme salinity , something that changes coat color of a rabbit, etc.

Tools for Recombinant DNA Technology

A Cloning Vector: they carry the Gene Of Interest (GOI) into the host organism. pUC19 used here is a cloning vector. Like other cloning vectors, pUC19 also have selectable markers. The 2 selectable markers in pUC19 are an ampicillin resistance gene (amp) and a LacZ gene (codes for enzyme beta-galactosidase).
Restriction Endonuclease: these are enzymes that enables us to make characteristic cuts in specific regions of DNA .First they help to cut out the GOI from their native organism. Then the same restriction enzyme is used to make similar cuts in the cloning vector . Due to the similarity in cuts (we call them as sticky ends) the GOI can be inserted into the vector. Vector containing the GOI is called a recombinant.
A competent host: The recombinant (vector containing GOI) is then incorporated into a host cell within which it will multiplicate and also get passed down into their progenies . Inside them the GOI gets expressed bringing about the desired result, which could be a biomolecule of our interest or some other trait.