You are investigating the activity of different cadherins in cells. You use recombinant DNA technology to express GFP-E-cadherin (green) in some L-cells (which do not normally contain cadherin) and YFP-P-cadherin (yellow) in other L-cells. Explain results you expect in three cultures containing: (1) only cells expressing GFP-E-cadherin, (2) only cells expressing YFP-P-cadherin, and (3) a mixture of GFP-E-cadherin-expressing and YFP-P-cadherin-expressing cells. Also predict how (a) knocking down the ability of the cells to synthesize fibronectin (FN) or (b) adding EGTA, which depletes free Ca2+ concentration in the culture medium, might affect the results.

You are investigating the activity of different cadherins in cells. You use recombinant DNA technology to express GFP-E-cadherin (green) in some L-cells (which do not normally contain cadherin) and YFP-P-cadherin (yellow) in other L-cells. Explain results you expect in three cultures containing: (1) only cells expressing GFP-E-cadherin, (2) only cells expressing YFP-P-cadherin, and (3) a mixture of GFP-E-cadherin-expressing and YFP-P-cadherin-expressing cells. Also predict how (a) knocking down the ability of the cells to synthesize fibronectin (FN) or (b) adding EGTA, which depletes free Ca2+ concentration in the culture medium, might affect the results.

Human Heredity: Principles and Issues (MindTap Course List)

11th Edition

ISBN:9781305251052

Author:Michael Cummings

Publisher:Michael Cummings

Chapter13: An Introduction To Genetic Technology

Section: Chapter Questions

Problem 20QP: Analyzing Cloned Sequences A base change (A to T) is the mutational event that created the mutant...

See similar textbooks

Similar questions

A research group is studying a bacterium X that binds to mucosal cells in the lung and invades. Wildtype X has an LD50 value of 10 bacteria when administered to mice by inhalation. Using transposon mutagenesis, the researchers have isolated two mutants of X that they call Xmut1 and Xmut2, both of which have LD50 values of 105 when inhaled by mice. However, in tissue culture cells, Xmut1 can invade the cells just as well as wild-type X, while Xmut2 cannot. Provide a possible explanation for these results.
T. Miyake and M. Demerec examined proline-requiring mutations in the bacterium Salmonella typhimurium (). On the basis of complementation testing, they found four proline auxotrophs: proA, proB, proC, and proD. To determine whether proA, proB, proC, and proD loci were located close together on the bacterial chromosome, they conducted a transduction experiment. Bacterial strains that were proC+ and had mutations at proA, proB, or proD were used as donors. The donors were infected with bacteriophages, and progeny phages were allowed to infect recipient bacteria with genotype proC− proA+ proB+ proD+. The recipient bacteria werethen plated on a selective medium that allowed only proC+ bacteria to grow. After this, the proC+ transductants were plated on selective media to reveal their genotypes at the other three pro loci. The following results were obtained: Q.Which genotypes represent single transductants and which represent cotransductants?
T. Miyake and M. Demerec examined proline-requiring mutations in the bacterium Salmonella typhimurium (). On the basis of complementation testing, they found four proline auxotrophs: proA, proB, proC, and proD. To determine whether proA, proB, proC, and proD loci were located close together on the bacterial chromosome, they conducted a transduction experiment. Bacterial strains that were proC+ and had mutations at proA, proB, or proD were used as donors. The donors were infected with bacteriophages, and progeny phages were allowed to infect recipient bacteria with genotype proC− proA+ proB+ proD+. The recipient bacteria werethen plated on a selective medium that allowed only proC+ bacteria to grow. After this, the proC+ transductants were plated on selective media to reveal their genotypes at the other three pro loci. The following results were obtained: Q.Is there evidence that proA, proB, and proD are located close to proC? Explain your answer.
T. Miyake and M. Demerec examined proline-requiring mutations in the bacterium Salmonella typhimurium (). On the basis of complementation testing, they found four proline auxotrophs: proA, proB, proC, and proD. To determine whether proA, proB, proC, and proD loci were located close together on the bacterial chromosome, they conducted a transduction experiment. Bacterial strains that were proC+ and had mutations at proA, proB, or proD were used as donors. The donors were infected with bacteriophages, and progeny phages were allowed to infect recipient bacteria with genotype proC− proA+ proB+ proD+. The recipient bacteria werethen plated on a selective medium that allowed only proC+ bacteria to grow. After this, the proC+ transductants were plated on selective media to reveal their genotypes at the other three pro loci. The following results were obtained: Q.Why are there no proC− genotypes among the transductants?
antibodies were produced in a rabbit against a 25 KDa human soluble protien. when the rabbit antiserum was used in a western blot of human soluble protiens, it detected 25-KDa protien,but also bound to protiens of 65KDa and 39 KDa.assuming that a pure protien was used to make the antibodies,how do yuo explain the results of the western blot?
After you have successfully cloned Sonic Hedgehog into pSELECT-CCFP-blasti and transfected HEK 293 cells (human embryogenic kidney cells), you observed, using confocal microscopy that the GFP fluorescence displays a reticulate network pattern in the cytoplasm of the cells. Describe an approach you can use to determine the subcellular localization of the Sonic Hedgehog-GFP fusion protein.
Can you please help me by drawing a serie of schematic figures that demonstrates the information in the paragraph below. The carboxy terminus of the p53 protein acts as an allosteric regulator of sequence-specific DNA binding. This was demonstrated initially by Hupp et al. (1992) using a bacterially expressed protein. Recombinant bacterial p53 bound poorly to DNA, and binding could be enhanced by the addition of antibodies specific to the C-terminal region of the protein. Phosphorylation of Ser315 and Ser392 within this domain also enhance sequence-specific DNA binding. Dephosphorylation of Ser376 of p53 after IR allows the association of 14-3-3 proteins with the C terminus of the protein (Waterman et al., 1998). Stavridi et al. (2001) demonstrate that this interaction is required for p53 to activate the downstream gene, p21waf1/cip1, and for the G1 cell cycle checkpoint arrest response. Interestingly, this dephosphorylation event seems to be ATM-dependent, possibly by a phosphatase…
Analyzing Cloned Sequences A base change (A to T) is the mutational event that created the mutant sickle cell anemia allele of beta globin. This mutation destroys an MstII restriction site normally present in the beta globin gene. This difference between the normal allele and the mutant allele can be detected with Southern blotting. Using a labeled beta globin gene as a probe, what differences would you expect to see for a Southern blot of the normal beta globin gene and the mutant sickle cell gene?
Regarding the production of recombinant monoclonal antibodies, it is CORRECT to state that: * (JUST ONE STATEMENT IS CORRECT) (A) Chimeric and humanized antibodies differ in relation to the percentage of sequence of the constant chain of molecules from the human genome (B) Chimeric antibodies are industrially produced by joining purified protein fragments of the Fab fraction of murine antibodies and the constant portion of human antibodies. (C) When using cells dependent on the DHF amplification system, we must use a vector containing the coding information of the light and heavy chains and another vector that contains the coding sequence of the enzyme dihydrofolate reductase (D)The industrial purification of monoclonal antibodies should preferably use an initial step based on the use of affinity columns, which will bind the antibodies by the Fc fraction (E) The use of amplification systems such as GS and DHFR has as main objective to make the production of monoclonal antibodies…
When Avery and his colleagues had obtained what was concluded to be the transforming factor from the IIIS virulent cells, they treated the fraction with proteases, RNase, and DNase, followed in each case by the assay for retention or loss of transforming ability. What were the purpose and results of these experiments? What conclusions were drawn?
Explain the process of how X-gal screening works with pUC19, you may build a model with boxes and arrows. 2. You are utilizing BamHI (GGATCC) restriction site and HindIII (AAGCTT) restriction site. Within pUC19, BamHI is at position 263, while HindIII is at position 233. (Hint: position is like coordinate on a map). If you manage to insert GTF2H5 in pUC19 vector, what are the sizes of fragments if you digest the pUC19-GTF2H5 (this is after insertion) with the following restriction enzyme combination after gel electrophoresis: BamHI and HindIII There is an NdeI site right in the middle of the GTF2H5 that has been inserted in pUC19, what would be the fragment sizes, if you digest with BamHI and NdeI. HindIII and NdeI BamHI, HindIII, and NdeI. 3. Design an experiment to confirm the presence of insert GTF2H5 in pUC19 vector using the following method (besides restriction analysis above), assuming that you know the sequence of GTF2H5: Southern Blot Polymerase Chain Reaction
For the experiment shown Fig. 6.30 and 6.31, what is the advantage to use a labeled incoming nucleotide (α-32P-UTP or α-32P-CTP), not labeled reagent 1 itself? Please draw a diagram to help explain your answer by contrasting the RNA polymerase labeled with α-32P-Reagent (Panel A) vs α-32P-UTP (Panel B).