2. In order to make a recombinant plasmid for transformation, EcoRI you digested a bacterial plasmid with restriction enzyme EcoRI. You want to insert a gene of interest that codes for 5... GAATTC...3' antibiotic resistance. You decide to try both XXRI and ZZRI 3... CTTAAG...5 restrictions enzymes for the antibiotic resistance gene. For XXRI, would you expect the gene insertion process to work? Why or why not? If you grow your transformed bacteria (assuming all other parts of a. XXRI 5... GAATTC...3 3... СТТАAG...5' transformation work), what would you expect to see on an LB/antibiotic plate? b. For ZZRI, would you expect the gene insertion process ZZRI to work? Why or why not? If you grow your transformed bacteria (assuming all other parts of 5... A A ATTT...3 3... TTTA AA... 5 transformation work), what would you expect to see on an LB/antibiotic plate?

2. In order to make a recombinant plasmid for transformation, EcoRI you digested a bacterial plasmid with restriction enzyme EcoRI. You want to insert a gene of interest that codes for 5... GAATTC...3' antibiotic resistance. You decide to try both XXRI and ZZRI 3... CTTAAG...5 restrictions enzymes for the antibiotic resistance gene. For XXRI, would you expect the gene insertion process to work? Why or why not? If you grow your transformed bacteria (assuming all other parts of a. XXRI 5... GAATTC...3 3... СТТАAG...5' transformation work), what would you expect to see on an LB/antibiotic plate? b. For ZZRI, would you expect the gene insertion process ZZRI to work? Why or why not? If you grow your transformed bacteria (assuming all other parts of 5... A A ATTT...3 3... TTTA AA... 5 transformation work), what would you expect to see on an LB/antibiotic plate?

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...

See similar textbooks

Similar questions

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.
Im designing a restriction cloning experiment. My professor said that plasmid and insert DNA work best in a 1:1 ratio. If my reaction does not work the first time I perform it, how would I ensure that the cause was the difference in concentration of reagents and how would I fix it so that the I have the correct concentration? If a restriction cloning experiment does not work, how could I be certain that it was due to an incorrect ratio of plasmid:insert and how can i fix this?
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)
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.
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
Which of the following would you need to clone a goat? Choose all that apply a) A restriction enzyme b) A surrogate mother c) A plasmid vector (or other type of DNA vector, e.g. phage, BAC) d) A donor goat nucleus e) Human DNA f) DNA ligase g) An enucleated egg
A plasmid has two restriction cut sites for EcoRI and one restriction cut site for HindIII. The plasmid is subjected to PCR, and the region containing the three restriction cut sites is amplified, such that all three sites are present in the resulting amplicon. If the amplicon is incubated with EcoRI, how many fragments of DNA are expected to result from this digest experiment? a. 0 b. 1 c. 2 d. 3 e. 4
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
You have ligated a foreign DNA fragment into the EcoRI site in multiple cloning site of pUCI18. You then transform how E.coli with the ligation mixture. Some of the bacterial colonies growing on the nutrient agar plate that contains ampicillin and Xgal are white and some are blue. Bacterial colonies that are white a) contain recombinant plasmid DNA b) have hydrolyzed ampicillin to form a colored compound c) are untransformed bacteria d) contain non recombinant plasmid DNA
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…