3. A plasmid was digested with the enzymes BglII, Nsil and XhoI. A single digest was performed with each enzyme and then double digests were performed. Make a restriction map of the plasmid based on the data below and indicate the distances between the restriction sites Plasmid cut BglII Nsil XhoINsil + Xhol Nsil + B9|II|B9|II + XhoI with: DNA| 33.2 31.2|47.8 fragments 18.2 20.2 3.6 (bp) 28.0 24.2 18.2 20.2 19.0 8.6 15.0 5.2 5.4 3.6 3.2 3.6

3. A plasmid was digested with the enzymes BglII, Nsil and XhoI. A single digest was performed with each enzyme and then double digests were performed. Make a restriction map of the plasmid based on the data below and indicate the distances between the restriction sites Plasmid cut BglII Nsil XhoINsil + Xhol Nsil + B9|II|B9|II + XhoI with: DNA| 33.2 31.2|47.8 fragments 18.2 20.2 3.6 (bp) 28.0 24.2 18.2 20.2 19.0 8.6 15.0 5.2 5.4 3.6 3.2 3.6

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 1TYK

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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.
Lane 5 in the gel above displays the pBashi cleavage products after restriction digest with the restriction enzymes Xhol+ Pstl. Lane 6 in the gel above displays the pBashi cleavage products after restriction digest with the restriction enzymes Xhol+ HindlII+ Pstl. label on the plasmid. i) The location of the Pstl site ii) The sizes of the new fragments made by this Pstl cut
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. Scientists were able to develop transgenic bacteria that can be used for mercury remediation. This process involved combining genes for mercury resistance (so that the bacteria itself has protection) and mercury accumulation (thereby allowing the bacteria to sequester Hg for retrieval later on). They probably used the following techniques in creating the transgenic bacteria EXCEPT a. plasmids b. DNA ligase c. DNA polymerase d. restriction enzymes 2. Consider these two parents (Mom and Dad) and their son, Calvin. (see pic attached). Which two members of this family would show the greatest difference in their DNA profile?
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
A student wanted to load .75 ug dna on agarose gel and has 4x loading buffer for sample preparation. The student has 50ul purified plasmid, finding concentration of plasmid to be 250 ng/uL The student used 10uL plasmid DNA in 50uL reaction for the restriction digest Give the volumes of restriction digest and 4x loading buffer that student would mix together and load on agarose gel.
1. Write if the statement is true and modify the statement with the correct answer if its false a.Blue colonies in the white-blue screening provide sufficient information on the transfer of recombinant DNA in bacterial cells. b.In blue-white screening, it is only the blue colonies that have survived the effect of antibiotics in the media. c.Bacterial transformation simply means the insertion of a recombinant plasmid into a bacterial cell. d.Competent cells are those bacterial cells that can accept the entry or re-entry of plasmids.
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. Given the following restriction endonucleases and the sequences of their corresponding restriction sites, which would be LEAST useful for cutting the plasmid and the foreign DNA to be inserted? [The arrows indicate where cuts are made.] (see img) a. EcoRI b. HindIII c. HaeIII d. BamHI 2. EcoRI and HindIII are two different restriction enzymes. If the DNA from two different sources were cut with either EcoRI or HindIII, which of the cut DNA fragments would NOT join together easily so that they could be sealed with ligase? a. E. coli DNA cut with EcoRI and human DNA cut with HindIII b. E. coli DNA cut with HindIII and mouse DNA cut with HindIII c. human DNA cut with EcoRI and chimpanzee DNA cut with EcoRI d. mouse liver DNA cut with HindIII and mouse kidney DNA cut with HindIII
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1. (a) Restriction sites are usually ______. Recombinant DNA Technology Restriction enzymes Ligase Palindromic sequences (b) Involves joining a donor DNA fragment of interest to a vector Recombinant DNA Technology Restriction enzymes Ligase Palindromic sequences
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)