In Biotechnology, gene cloning is a very important technique. A vector is normally required to perform this process. The vector commonly used to transform a bacterial host cell is the plasmid. (i) State the THREE (3) important regions of the plasmid. Elaborate your answer.
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- In Biotechnology, gene cloning is a very important technique. A vector is normally required to perform this process. The vector commonly used to transform a bacterial host cell is the plasmid. (i) State the THREE (3) important regions of the plasmid. Elaborate your answer. (ii) Besides plasmids, name TWO (2) other commonly used vectors in Biotechnology.Cloning vectors are not just limited to bacterial plasmids. Bacteriophages and M13 phage vectors are also commonly utilized in the cloning process. State any five (5) key criteria to be an effective cloning vector.With the use of well-illustrated diagrams, reconstruct the entire cloning process by explaining different stages of the cloning process that involves the following:a. Isolation of target DNA fragments (often referred to as inserts)b. Ligation of inserts into the plasmid, creating recombinant molecules c. Transformation of recombinant plasmids into bacteria or other suitable host for propagationd. Screening/selection of hosts containing the intended recombinant plasmid. For this stage(d), discuss the importance of a second marker that can be used for screening of genomic DNA for colonies containing the pka-1 under the principle of insertional inactivation. This should be properly explained using all the attributes of the plasmid described above.
- A) Outline the experimental procedure for cloning a eukaryotic gene and expressing it in E. coli. Focus on the essential steps starting with eukaryotic gene amplification to transformation of E. coli cells B) Explain how insertional inactivation can help you identify the colonies that carry the plasmid with your eukaryotic gene of interest C) Plasmids containing antibiotic resistance genes are widely used in gene cloning and other molecular biology techniques. What would happen if the eukaryotic gene was inserted into an antibiotic resistance gene on the plasmid?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?There may be more than one correct answer, select all that apply. To clone genes, a plasmid vector should contain a(n) a.) telomeres b.) orgin of replication c.) antibiotic resistance gene d.) restriction endonuclease cut site The CRISPR/Cas system has revolutionized the efficiency at which scientists can experimentally induce a mutation in a gene to investigate the functional role of the protein it encodes. Which of the following are required in the CRISPR/Cas system? Cas9 nuclease Bacterial genome Access to target gene Guide RNA complementary to gene of interest
- You discovered a new gene and cloned it into a plasmid vector. You have reason to suspect this gene is a proto oncogene. Explain how you can test your hypothesis to experimentally determine if the new gene is a proto oncogene. Be specific about the types of biological array/blot or other machinery or technoques used.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…What type of enzymes are used to “cut” desired DNA sequences for use in recombinant gene technology experiments? Identify those two enzymes used to cut and paste both genes into the plasmid. Identify all three strategies used in this lab to maximize transformation success. Explain what it means for bacteria to be “competent.” Explains why bacterial competency is this important for this investigation.
- In the following "gene library" cloning experiment. Digested genomic DNA AmpR gene TCR gene TCR is tetracycline resistant marker, AmpR is ampicillin resistant marker and BamHI is the unique restriction enzyme on plasmid. Cells transformed with the "recombinant plasmids" will be:....... (Hint: this is a challenging question) a) resistant to ampicillin and tetracycline b) sensitive to tetracycline and ampicillin c) resistant to tetracycline and sensitive to ampicillin d) resistant to ampicillin and sensitive to tetracycline e) sensitive to ampicillin and tetracycline BamHIThe plasmids from the pUC series are created in the University of California. They carry a lacz gene that plays a significant role in the screening process after transformation. (i) Name the screening process utilizing the lacZ gene. (ii) Elaborate how this gene plays a crucial role in the screening step as mentioned above.Jackson Wang is a biologist working with the genetics of a thermophilic bacterium. He cloned a heat shock gene from the bacteria for further analysis. After cloning, he isolated the plasmid carrying his gene of interest for sequencing. Jackson finally received the nucleotide sequence of his gene. Explain in detail how he could verify whether the nucleotide sequence matches his gene of interest using the bioinformatics databases available.