. As described in the textbook, the plasmid vector pBluescript II contains an ampicillin resistance gene and a lacZ gene, within which is located a multiple cloning site. Briefly describe how one selects for/identifies bacterial cells containing recombinant pBluescript II molecules and explain how the selection/identification works.

. As described in the textbook, the plasmid vector pBluescript II contains an ampicillin resistance gene and a lacZ gene, within which is located a multiple cloning site. Briefly describe how one selects for/identifies bacterial cells containing recombinant pBluescript II molecules and explain how the selection/identification works.

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

Chapter17: Bacterial And Viral Genetics

Section: Chapter Questions

Problem 1TYK

See similar textbooks

Similar questions

1 (a) What do ApR, TcR, and ori on the pBR322 map represent and discuss there individual functions? (b)Does the undigested plasmid show more than a single band when electrophoresed? Why? (c)What other kinds of molecules, in addition to plasmid DNA would you expect to find in a sample of plasmid DNA extraction? (This is not a graded assignment)
1. Write if the statement is true and modify the statement with the correct answer if its false a. E. coli strains used in blue-white screening produce blue color complexes because their beta-galactosidases are activated via alpha-complementation. b. If you are a researcher trying to identify a gene that is responsible for a trait of interest, the first thing that you would do is to make a review of the literature that is available. c. The polylinker region in plasmids is where specific DNA sequences are cut by non-specific restriction enzymes. d. Blue-white screening is one of the last steps involved in determining if the gene of interest has been inserted successfully in plasmids.
1. Write if the statement is true and modify the statement with the correct answer if its false a.White colonies represent those bacteria that do not express the genes for beta-galactosidase because restriction enzymes have cut the gene and ligase have not repaired the break b.Antibiotic resistance genes in bacterial transformation are used as marker genes because of ease of detection of transformants. c. Bacterial transformation simply means the insertion of a recombinant plasmid into a bacterial cell.
1. Briefly explain why the total size of the pMBBS plasmid in the Restriction Enzymes practical is 3000bp (base pairs) 2. Briefly explain why the cut sites on the pMBBS plasmid in each Restriction Enzymes (EcoRI, BamHI, and XhoI) just 1 each 3. Briefly explain what led to the 5 fragments formed which linked to the 500bp, 1000bp, 1500bp, 2000bp, 2500bp sizes
1. How would you insert your gene of interest in a target location using CRISPR/Cas9? a. Introduce a plasmid containing the DNA sequence of gene of interest flanked by direct repeats. b. Introduce a plasmid containing the DNA sequence of gene of interest flanked by sequences similar to both sides of the target cut site. c. Introduce a plasmid containing the gRNA of your gene of interest d. Introduce a plasmid containing the DNA sequence of gene of interest flanked by inverted repeats.
1. Below is an illustration of two plasmids: the incomplete pKAN-R which contains the gene of interest (rfp), and the good plasmid vector pARA which contains the ampR gene for ampicillin resistance, the araC activator gene and the ori region. Based on this, how are you going assemble to create the desired recombinant plasmid containing all the necessary components? (Keywords: Identify correct restriction enzymes to use to create specific sticky end sequences that will only assemble with the final vector in one orientation)
With the use of well-illustrated diagrams, reconstruct the entire cloning process by explaining different stages of the cloning process that involves the following: d. 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.
1. Match the features of plasmids below with their meaning : Origin of replication LacZ gene antibiotic resistance gene Multiple cloning site A.required for maintenance of the plasmid in E. coli cells used to select for coli that contain our plasmid B. Required for replication of plasmid DNA inside an E. coli cell C. An engineered region of the plasmid that contains many unique restriction enzyme recognition Sites D. Allows for screening for plasmids with an insert in the multiple cloning site
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.
1. What two restriction sites are you going to use to clone your PCR product into the pL4440 plasmid? What are their DNA sequence? 2. State the primer sequence you will use to amplify the F27C1.7 gene ready to be cloned into the pL4440 plasmid? 3. How would you go about cloning this amplified DNA into pL4440? Using your knowledge of cloning list 5 important aspects of the method.
1. It is a double-stranded , self replicating, circular DNA molecule present in bacteria which is widely used as a gene cloning vector.A. Cosmid B. Plasmid. C. Phagemid. D. Genome 2. In plant genetic engineering, which of the following acts as vector?A. Agrobacterium lumefaciens. B. Gene of interest C Receipt plant cell. D. Ti-plasmid 3. Which of the following illustrates breeding?A. A farmer choose a breed of cow for greater milk productionB. The use of bacteria in order to produce human insulin.C. The insertion of cloned genes to plant cells.D. All of the above. 4. Being pest resistance is one of the traits that are being introduced to plants like corn and eggplant with the insertion of Bt-toxin gene to plant cell. What method is use when a gene is inserted to plants?A. Biolistic B. Electroporation C. Plasmid insertion by heat shock treatment D. All of these 5. A biotechnologist wants to enhance the size of duhat fruit because research shows that it can lower blood sugar but is very…
1)Find a plasmid map for pET11a and create a basic procedure for cloning a gene into this vector. Which selection method and substance would you use for this plasmid after transformation? 2) Which organism would be your first choice for transformation: a bacterium, earthworm, fish or mouse? Describe your reasoning. 3) What is the main difference between plates before and after centrifugation? What may be the purpose of concentrating the media and perform a second plating?