4. Consider that you identified an enzyme in a fungus which can digest plastic. You want to isolate and express this enzyme in E.Coli bacteria in high amounts. Design the main steps of cloning experiment. Explain how you will prepare the recombinant plasmid and how you will select the transformed bacteria colonies with recombinant plasmid.

4. Consider that you identified an enzyme in a fungus which can digest plastic. You want to isolate and express this enzyme in E.Coli bacteria in high amounts. Design the main steps of cloning experiment. Explain how you will prepare the recombinant plasmid and how you will select the transformed bacteria colonies with recombinant plasmid.

Biology: The Dynamic Science (MindTap Course List)

4th Edition

ISBN:9781305389892

Author:Peter J. Russell, Paul E. Hertz, Beverly McMillan

Publisher:Peter J. Russell, Paul E. Hertz, Beverly McMillan

Chapter18: Dna Technologies: Making And Using Genetically Altered Organisms, And Other Applications

Section: Chapter Questions

Problem 3TYK: Why are antibiotic resistance markers such as ampR important components of bacterial plasmid cloning...

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In bacterial transformation, the purpose of having antibiotic within an agar plate is to: Select one: confirm which plasmids been have successfully ligated with a gene of interest. isolate bacteria which have been successfully transformed with the plasmid. indicate which plasmids were successfully digested by the endonuclease. act as a substrate which will be cleaved and produce a blue product when ligation is unsuccessful. show which plasmids contain the lacZ gene.
35) A researcher inserted DNA fragments from an organism into expression vector plasmids and introduced the modified plasmids into bacterial cells. Which of the following methods would be an effective means of identifying which clones contain a specific gene of interest?
Second part asks: Explain the roles two of those enzymes would play in obtaining the recombinant plasmids
fomP is responsible for the chemical transformation of microplastics into ultra-efficient insulation. You take an arctic seawater sample and extract the DNA. 1. First you need to locate the gene on the bacterial chromosome. What procedure(s) would you use to identify and locate the gene? Explain how it/theywork(s). 2. Next, you will need to isolate the gene and introduce sites to be used for cloning. What would you use to make many copies of this gene? What will you need? How does it work on a molecular level?
As this is a non-directional cloning, recombinant plasmids can contain an insert ligated into the vector in two different orientations. Provide 2 diagrams to illustrate the 2 potential recombinant plasmids, with the inserts ligated in opposite orientations. Include all RE sites and distances between sites on the diagram.
1) Given the image below, describe what is happening at each of the 3 sections (i.e. 1, 2, and 3) . [In section 1, the gene donor is providing..., In section 2..., In section 3...]
3A. Following transformation with the CRISPR plasmid, the yeast will be plated on SD-URA plates. Why is SD-URA media used? Name the relevant marker gene in the pCRCT CRISPR plasmid. 3B. What would you expect to see if you plated on YED accidentally? Will most of the yeast be red or white? Why? 3C. The complete genotype of the host yeast strain is MATa ade2 his3 leu2 met15 ura3. Name one advantage of using a yeast strain with auxotrophies for several genes (i.e. his3, leu2 and ura3). 3D. After successful CRIPSR editing, the yeast can later be “cured” of the recombinant CRISPR plasmid – that is, the plasmid is lost but the CRISPR edit is stably inherited. Write the new genotype of the yeast-based on the ADE6 gene.
You have set up a recombinant DNA experiment using the plasmid PBR322 as the vector (see plasmid below). You use the BamHI restriction site on the plasmid to insert the target DNA. The plasmid is then used to transform E.coli colls Is the following statement True or False? Growth of the transformed cells on agar containing both ampicillin and tetracycline will eliminate any cells that do not contain a plasmid. Clal Hindlll EcoRI Pvul BamHI Pstl amp tet PBR322 -Sall ori rop Pvull True False
Using the Figure below briefly describe how the amplification can be used in the cloning process of DNA.
Please help with this as soon as you can, thank you.
dont provide handwriting solution ...
A.) Transformation is best described as: Group of answer choices The integration of foreign DNA into a genome The uptake of naked DNA from the environment The transfer of DNA via a bacteriophage Transfer of a plasmid from one organism to another B.) What is the function of the araC gene in the pGLO plasmid? Group of answer choices It encodes the protein that glows under ultraviolet light It allows us to select for the cells that contain the plasmid It prevents the transcription of the green fluorescent protein unless arabinose is added It ensures that the plasmid will be copied and passed on to daughter cells C.) What is the function of the bla gene in the pGLO plasmid? Group of answer choices It encodes the protein that glows under ultraviolet light It allows us to select for the cells that contain the plasmid It prevents the transcription of the green fluorescent protein unless arabinose is added It ensures that the plasmid will be copied and passed on…