Please put in correct order You are cloning a gene from a eukaryotic organism into a plasmid to produce a recombinant protein in bacteria. Put the following steps in the correct orderScreen the colonies that contain the right constructGrow bacteria for the expression of the gene you clonedInsert the gene into the plasmid by ligation reactionPlate the transformation reactionDigest the plasmid with the same restriction enzymesRun an agarose gell and isolate the DNA fragmentsSynthesize CDNATransform the bacteria with the plasmid you constructedIsolate the mRNACreate restriction sites by PCRP Type here to search

The biological process by which naked DNA (deoxyribonucleic acid) gets transferred from one…

Answered: You are cloning a gene from a…

You are cloning a gene from a eukaryotic organism into a plasmid to produce a recombinant protein in bacteria. Put the following steps in the correct order

Human Heredity: Principles and Issues (MindTap Course List)

11th Edition

ISBN:9781305251052

Author:Michael Cummings

Publisher:Michael Cummings

Chapter13: An Introduction To Genetic Technology

Section: Chapter Questions

Problem 8QP

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Cloning Genes Is a Multistep Process In cloning human DNA, why is it necessary to insert the DNA into a vector such as a bacterial plasmid?
Which of the following best describes the process of DNA sequencing? a. DNA is separated on a gel, and the different bands are labeled with fluorescent nucleotides and scanned with a laser. b. A laser is used to fluorescently label the nucleotides present within the DNA, the DNA is run on a gel, and then the DNA is broken into fragments. c. Nucleotides are scanned with a laser and incorporated into the DNA that has been separated on a gel, and then the DNA is amplified with PCR. d. Fragments of DNA are produced in a reaction that labels them with any of four different fluorescent dyes, and the fragments then are run on a gel and scanned with a laser. e. DNA is broken down into its constituent nucleotides, and the nucleotides are then run on a gel and purified with a laser.
Figure 17.7 You are working in a molecular biology lab and, unbeknownst to you, your lab partner left the foreign genomic DNA that you are planning to clone on the lab bench overnight instead of storing it in the freezer. As a result, it was degraded by nucleases, but still used in the experiment. The plasmid, on the other hand, is fine. What results would you expect from your molecular cloning experiment? There will be no colonies on the bacterial plate. There will be blue colonies only. There will be blue and white colonies. The will be white colonies only.
A biologist is attempting to clone the gene encoding a particular enzyme (Enz) into a plasmid vector in E.coli. This plasmid has a gene encoding a green fluorescent protein (GFP) as well as a gene for tetracycline antibiotic resistance (TetR). The restriction site (to clone foreign DNA into) is within the GFP sequence. Which of the following would be expected when trying to see which E. coli cells acquired the recombinant plasmid (i.e., carrying the Enz gene)? Bacteria UNABLE to grow on tetracycline-containing media AND are NOT able to make green fluorescent protein are the ones that contain the recombinant plasmid. Bacteria able to grow on tetracycline-containing media AND that are NOT able to make green fluorescent protein are the ones that contain the recombinant plasmid. Bacteria able to grow on tetracycline-containing media AND are able to make green fluorescent protein are the ones that contain the recombinant plasmid. Bacteria UNABLE to…
Now that you’ve isolated the gene and made lots of copies, you need to insert the gene into something you can manipulate and move into your lab strain of E. coli. You have access to the following vectors, either pBR322 or pUC19 (look them up here to see a map https://www.neb.com/products/dna-plasmids-and-substrates Please note the restriction sites in BOLD appear only once in the plasmid). You may use either vector. What is a vector? Which vector did you choose? Explain why you made that choice. What enzyme(s) will you use to place your fragment into the vector? Explain why you chose these enzymes. How will you move this construct into coli? What phenotype will tell you if the coli have taken up the plasmid? What phenotype will tell you if the coli have taken up the plasmid with the gasP gene?
A plasmid vector has two genes in it, a gene for streptomycin resistance (strR) and another for kanamycin resistance (kanR). There is a restriction site inside of the strR gene that you will use to move in your gene of interest. After cutting both your vector and insert you mix them together in a test tube. You then transform your mixture into E. coli and plate your cells on a master plate without any antibiotics. After colonies appear, you pick and re-plate onto a streptomycin containing plate and onto a kanamycin containing plate. What do you expect to grow on the master plate without any antibiotics? What do you expect to grow on the streptomycin plate? What do you expect to grow on the kanamycin plate? How do you determine which colonies contain your gene of interest? As mentioned above, there is a restriction site found within the strR Please draw or write out two different examples of what restriction sites may look like. These should be short…
A plasmid vector has two genes in it, a gene for streptomycin resistance (strR) and another for kanamycin resistance (kanR). There is a restriction site within the strR gene that you are hoping to move your gene of interest into. After cutting both your vector and insert you mix them together in a test tube. You then transform your mixture into E. coli and plate your cells on a master plate without any antibiotics. After colonies appear, you pick and re-plate onto a streptomycin containing plate and onto a kanamycin containing plate. 1. Using those same pieces of short DNA that you created above, show me what one of them may look like if it was cut with a blunt end restriction endonuclease. With the other example, show me what it would look like if it was cut with a sticky end restriction endonuclease. 2. Why do restriction endonucleases exist in bacterial cells? What must they do to protect their own DNA?
A plasmid vector has two genes in it, a gene for streptomycin resistance (strR) and another for kanamycin resistance (kanR). There is a restriction site within the strR gene that you are hoping to move your gene of interest into. After cutting both your vector and insert you mix them together in a test tube. You then transform your mixture into E. coli and plate your cells on a master plate without any antibiotics. After colonies appear, you pick and re-plate onto a streptomycin containing plate and onto a kanamycin containing plate. 1. What do you expect to grow on the master plate without any antibiotics? 2. What do you expect to grow on the streptomycin plate? 3. What do you expect to grow on the kanamycin plate?
A plasmid vector has two genes in it, a gene for streptomycin resistance (strR) and another for kanamycin resistance (kanR). There is a restriction site within the strR gene that you are hoping to move your gene of interest into. After cutting both your vector and insert you mix them together in a test tube. You then transform your mixture into E. coli and plate your cells on a master plate without any antibiotics. After colonies appear, you pick and re-plate onto a streptomycin containing plate and onto a kanamycin containing plate. 1. How will you determine which colonies contain your gene of interest? Be specific about what you will be screening for. 2. As mentioned above, there is a restriction site found within the strR gene. Please draw or write out two different examples of what restriction sites may look like. These should be short pieces of double stranded DNA.
Which statement is NOT the function of restriction enzyme? Group of answer choices It cuts the vector’s plasmid and the desired gene from the original DNA. It ligates the desired to gene to vector’s plasmid. It acts as scissors of DNA that cut along a specific sequence. It serves as degrading proteins that cut plasmid on specific target site.
You want to clone a specific PCR amplicon. You have determined that the amplicon you want to clone has enzyme restriction sites for HindIII and EcoRI. After investigation you have seen that the pUC18/19 plasmid also have these enzyme restriction sites in its multiple cloning site (MCS on map included). After enzyme digestion your amplicon is 854 bp long. What length will the recombinant plasmid be after you have inserted your amplicon? Show a calculation.
Below is shown a plasmid map from a recombinant DNA experiment you have performed. You inserted your gene of interest (labeled gene on the map) into the lac Z reporter gene via a Hind III site. If you digested this recombinant DNA with HindIII what is the size of the DNA fragment(s) you would obtain on a DNA gel electrophoresis? The plasmid alone is 4000 bp. I have attached the image below.