2. Your lab mate, Beaker, tells you that they have run into some issues with expressing a gene on a plasmid they've recently created. The gene has a proper promoter and a lacO sequence and no lacl sequence on the plasmid so that there will be constitutive expression of the gene. Beaker used standard protocols to transform wild-type E. coli with this plasmid and verified the colony they chose has the plasmid with the gene. However, Beaker cannot seem to recover any gene product (MRNA or protein) from a standard culture with appropriate media. What has your lab mate done wrong with this experiment? . What options are there for your lab mate to fix this issue? There are three possible options, specify two options and justify them.

2. Your lab mate, Beaker, tells you that they have run into some issues with expressing a gene on a plasmid they've recently created. The gene has a proper promoter and a lacO sequence and no lacl sequence on the plasmid so that there will be constitutive expression of the gene. Beaker used standard protocols to transform wild-type E. coli with this plasmid and verified the colony they chose has the plasmid with the gene. However, Beaker cannot seem to recover any gene product (MRNA or protein) from a standard culture with appropriate media. What has your lab mate done wrong with this experiment? . What options are there for your lab mate to fix this issue? There are three possible options, specify two options and justify them.

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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Similar questions

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. 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
2. You are cloning the genome of a new DNA virus into pUC18. You plate out your transformants on ampicillin plates con-taining X-gal and pick one blue colony and one white colony. When you check the size of the inserts in each plasmid (blueand white), you are surprised to fi nd that the plasmid fromthe blue colony contains a very small insert of approximately60 bp, while the plasmid from the white colony does notappear to contain any insert at all. Explain these results.
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 (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. You are having problems getting good transfection efficiency when you put your plasmid in e. coli. You want to freeze down a stock of e. coli that is more than 99% transfected. What modifications can you make to your plasmid and what steps can you take to ensure that 99% of your e. coli are transfected?
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)
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.
You isolated a 10,500 base pair plasmid (supercoiled, cccDNA) from E. coli. The plasmid contains one unique recognition site for EcoRI, a restriction enzyme. Restriction enzymes recognize a specific sequence and cut both strands of the DNA at that sequence (Chapter 7). Brief DNase I treatment breaks one (or very few) phosphodiesterbonds in each DNA molecule, leaving the double-stranded DNA with one strand broken but the other strand intact, i.e “nicked.” You briefly incubated the cccDNA at 37°C in four separate reactions containing the components listed below. You ran each reaction sample on an agarose gel and visualized the DNA using ethidium bromide and UV light. The reactions included the appropriate buffer and ATP when required. An agarose gel containing four lanes of possible products is given below. Please refer to Figs. 4-26 and 4-27 in Watson for reference. For each reaction, indicate which lane of the gel contains the products that you would expect to see on your…
ECOR1 cuts Plasmid T into 3 pieces. Feeling grumpy at your professor (☹), you decide to take a random enzyme and mix it with a random plasmid, Plasmid M (for mystery). You run a gel and see one very large band at the top of the gel. Which of the following is NOT likely: a. Plasmid M has zero restriction sites for any enzyme b. The enzyme and plasmid are not the same as Ecor1 and Plasmid T c. Plasmid M wasn't cut at all d. The plasmid does not have a restriction site for the mystery enzyme Please answer asap and type your answer and do not copy from anywhere please
1. How may recombinant DNA molecules be introduced into human cells? a. by splicing the needed genes into a mammalian chromosome using restriction enzymes. b. by adding plasmids to the mammalian cells. c. by using engineered viruses as vectors. d. by using a gene gun. 2. If someone is accused of a crime, investigators can collect his or her DNA to compare the DNA of the cells found at the crime scene. To collect human DNA, investigators often swab the inside of person’s cheek. Just a few human cheek cells contain enough material to perform PCR. In a cell, the nucleus and mitochondria contain DNA that is the starting material for PCR. Identify the 4 components needed to start a PCR reaction (equipment not included)
1. What the most important components a recombinant plasmid must have? (Keywords: gene of interest/target gene, promoter region, ori/origin of replication, marker/resistant gene)