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Molecular Cell Biology Lodish 7th Edition Test Bank

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9 Visualizing, Fractionating,
and Culturing Cells

PART A: Linking Concepts and Facts

9.1 Organelles of the Eukaryotic Cell

1. Which of the following synthesizes the phospholipids that form the plasma membrane?
a. the rough endoplasmic reticulum
b. the plasma membrane
c. the Golgi apparatus
d. the smooth endoplasmic reticulum

Ans: d

2. The pH of lysosomes is lower than that of the cytosol because of the action of
a. Na+ and OH− transport proteins in the lysosomal membrane.
b. H+ and Cl− transport proteins in the plasma membrane.
c. acid-producing enzymes in the lysosomal lumen.
d. H+ and Cl− transport proteins in the lysosomal membrane.

Ans: d

3. The oxidation of fatty acids occurs in the

a. mitochondria.
b. rough endoplasmic reticulum.
c. peroxisome.
d. a and c

Ans: d

4. __________ is (are) composed of a series of compartments that function to sequentially modify proteins and
lipids.
a. The endoplasmic reticulum
b. The Golgi apparatus
c. Peroxisomes
d. Vacuoles

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 9-2

Ans: b

9.2 Microscopy: Visualizing Cell Structure and Localizing Proteins Within Cells

5. The phenomenon in which a chemical absorbs light at one wavelength and emits it at a specific and longer
wavelength is called
a. differential interference contrast.
b. fluorescence.
c. deconvolution.
d. shadowing.

Ans: b

6. Which of the following could be used to visualize subcellular structure in living cells?
a. transmission electron microscopy
b. scanning electron microscopy
c. brightfield microscopy
d. differential interference light microscopy
Ans: d

7. Osmium tetroxide is commonly used to

a. stain specimens for light microscopy.
b. coat specimens for metal shadowing electron microscopy.
c. stain specimens for transmission electron microscopy.
d. measure the Ca2+ concentration inside living cells.

Ans: c

8. To visualize cells by immunofluorescence microscopy, the cells must be

a. placed in a vacuum.
b. living.
c. sectioned.
d. permeabilized.

Ans: d

9. The fluorescent properties of dyes such as SNARF-1 can provide information on the
a. location of specific proteins.
b. concentration of H+ ions in specific regions of the cell.
c. the amount of RNA in a cell.
d. volume of a cell.

Ans: b

10. Which of the following allows one to circumvent the theoretical resolution of the microscope?
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a. total internal reflection fluorescence microcopy

b. photo-activated localization microscopy
c. indirect immunofluorescence microscopy
d. double-label fluorescence microscopy

Ans: b

9.4 Purification of Cell Organelles

11. If a cellular homogenate were subjected to differential centrifugation, which of the following would be expected
to pellet first?
a. the endoplasmic reticulum
b. mitochondria
c. the cytosol
d. nuclei

Ans: d

12. The disruption of a cell is necessary to release its organelles and contents for subsequent isolation. One method,
called _________________________________, uses ultrahigh-frequency sound to disrupt the cell plasma
membrane.
a. tomography
b. epitope tagging
c. sonication
d. centrifugation

Ans: c

13. Although many types of vesicles are similar in size and density, it is possible to isolate specific types of vesicles
through the use of
a. a fluorescent-activated cell sorting machine.
b. antibodies attached to bacterial carriers and low speed centrifugation.
c. ultracentrifugation.
d. light microscopy.

Ans: b

14. Ultracentrifuges allow cell biologists to isolate mitochondria from lysosomes based on organelle differences in

a. isoelectric point.
b. ionic composition.
c. equilibrium density.
d. size.

Ans: c
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9.5 Isolation, Culture, and Differentiation of Metazoan Cells

15. A myeolma cell is best described as

a. a precursor cell that gives rise to gametes.

b. an immortal immune cell that cannot produce antibodies.
c. a self-renewing stem cell.
d. a and c

Ans: b

16. What factors necessary for growth of animal cells in culture are provided by serum?
a. amino acids
b. precursors of DNA synthesis
c. growth factors
d. vitamins

Ans: c

17. Which one of the following is the best technique/approach to allow you to localize catalase in peroxisomes?

a. a catalase monoclonal antibody and transmission electron microscopy

b. platinum or gold and scanning electron microscopy
c. FRAP and FRET
d. all of the above

Ans: a

18. All of the following are produced by animal cells in culture and help the cells adhere to the culture dish except
a. glycoproteins.
b. collagen.
c. phospholipase A.
d. hyaluronic acid.

Ans: c

19. Characteristics of transformed cells can include all of the following except
a. aneuploidy.
b. ability to differentiate.
c. tight junctions.
d. presence of integrated viral genes.

Ans: c
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PART B: Testing on the Concepts

9.1 Organelles of the Eukaryotic Cell
20. How do integral proteins travel from the ER to the Golgi and from the Golgi to the plasma membrane?
Ans: Integral proteins travel in the membrane of transport vesicles that move from the ER to the Golgi and from the
Golgi to the plasma membrane.

21. Each organelle has a specific function in the eukaryotic cell. What determines the function of each organelle?

Ans: The specific function of a given organelle is determined by the specific proteins present in that organelle.

9.2 Light Microscopy: Visualizing Cell Structure and Localizing Proteins Within Cells
22. How does the wavelength of the light used to illuminate a specimen affect the ability to resolve objects within
the specimen?
Ans: Because the limit of resolution is given by D = (0.61/(N sin ), shorter wavelength light (e.g., blue) will
provide better resolution than longer wavelength light (e.g., red).

23. Fixatives such as formaldehyde are routinely used in certain types of electron microscopy and light
microscopy. However, fixatives may introduce complications in analysis of the resulting images. What
problems may result from using fixatives?

Ans: Most fixatives are cross-linking agents that immobilize proteins and other cellular molecules. As part of the
fixation process, samples are often also dehydrated. Such chemical changes may alter the normal structure and
spatial relationship of cellular components, and this must be taken into account when interpreting images of
fixed specimens.

24. You are studying lamins and have an antibody method to follow its expression and subcellular localization at
the electron microscope level. Where would you find these proteins in the bacteria E. coli?

Ans: Normally, antibody labeling can be used in tandem with electron microscopy to visualize proteins in thin
sections. Cells are lightly fixed to avoid denaturing the epitopes on the desired protein and, after freezing, the
tissue is sectioned at very low temperature. The tissue is thawed and a specific antibody to the desired protein
is applied as in immunofluorescence microscopy. To detect the antibody, however, the tissue is incubated with
electron-dense gold particles attached to protein A, a bacterial protein that binds the Fc segment of all
antibodies. These gold particles are seen at the level of the electron microscope, however, you would not be
able to see lamins in E. coli because bacteria do not contain membrane-bounded organelles and lamins are a
component of the nuclear envelope.

25. Describe how a Förster resonance energy transfer biosensor like cameleon, which consists of CFP linked to YFP
by the protein calmodulin, can detect local changes in calcium ion concentration. For your information, CFP
excites at 440 nm and emits at 480 nm, whereas YFP emits at 535 nm.

Ans: Basically, if calcium is absent and the cameleon construct is excited by UV light at 440 nm, there is emission
of energy at 480 nm, but because the distance between CFP and YFP is greater than 10 nm, the energy cannot
Molecular Cell Biology Lodish 7th Edition Test Bank

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be transferred to YFP and hence there is no detection of light at 535 nm. In the presence of calcium ions,
however, calmodulin undergoes a conformational change, which brings CFP in close proximity to YFP. When
CFP is excited with UV light at 440 nm, the energy is transferred to YFP, allowing it to fluoresce and be
detected at 535 nm.

9.4 Purification of Cell Organelles

26. Rough endoplasmic reticulum can be separated from smooth endoplasmic reticulum by differential
centrifugation. What is the basis for this fractionation?

Ans: During cell disruption, the endoplasmic reticulum fragments and reseals into small vesicle-like compartments
termed microsomes. Microsomes derived from the rough endoplasmic reticulum contain ribosomes and these
ribosomes provide additional mass, which allows separation from microsomes derived from the smooth
endoplasmic reticulum.

9.5 Isolation, Culture, and Differentiation of Metazoan Cells

27. Separation of most blood cells is difficult, if not impossible, to achieve because they have similar properties
and/or densities. What procedure is used to separate T-cells of the immune system from the many other
different types of white blood cells or spleen cells? What feature of the T-cell facilitates the isolation
protocol?

Ans: Flow cytometry and fluorescence-activated cell sorting is used to select and isolate T cells away from the
excess number of other cell types. Briefly, T cells, unlike other cells, express CD3 and Thy1.2 proteins on
their cell surface. Antibodies specific to these markers are linked to a fluorescent dye and incubated with the
pool of cells. The antibodies bind the cell-surface markers on the T-cell surface and when all cells are placed
in the FACS machine, a laser is used to excite the dye causing it to fluoresce. Fluorescing T cells, selectively
sorted from the non-fluorescing cells, can be cultured in vitro.

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