Name: __________________________ Date: _____________
1. Using terms from the list below, fill in the blanks in the following brief description of
the experiment with Streptococcus pneumoniae that identified which biological
molecule carries heritable genetic information. Some terms may be used more than
once.
Cell-free extracts from S-strain cells of S. pneumoniae were fractionated to
__________________ DNA, RNA, protein, and other cell components. Each fraction
was then mixed with __________________ cells of S. pneumoniae. Its ability to change
these into cells with __________________ properties resembling the
__________________ cells was tested by injecting the mixture into mice. Only the
fraction containing __________________ was able to __________________ the
__________________ cells to __________________ (or __________________ ) cells
that could kill mice.
carbohydrate lipid R-strain
DNA nonpathogenic RNA
identify pathogenic S-strain
label purify transform
2. The structures of the four bases in DNA are given in Figure Q5-20.
Figure Q5-20
A. Which are purines and which are pyrimidines?
B. Which bases pair with each other in double-stranded DNA?
3. Given the sequence of one strand of a DNA helix (below), provide the sequence of the
complementary strand and label the 5 and 3 ends.
5-GCATTCGTGGGTAG-3
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4. When double-stranded DNA is heated, the two strands separate into single strands in a
process called melting or denaturation. The temperature at which half of the duplex
DNA molecules are intact and half have melted is defined as the Tm.
A. Do you think Tm is a constant, or can it depend on other small molecules in the
solution? Do you think high salt concentrations increase, decrease, or have no effect on
Tm?
B. Under standard conditions, the expected melting temperature in degrees Celsius
can be calculated from the equation Tm = 59.9 + [0.41 × %(G + C)] – (675/length of
duplex). Does the Tm increase or decrease if there are more G + C (and thus fewer A +
T) base pairs? Does the Tm increase or decrease as the length of DNA increases? Why?
C. Calculate the predicted Tm for a stretch of double helix that is 100 nucleotides
long and contains 50% G + C content.
5. A. Define a gene.
B. Consider two different species of yeast that have similar genome sizes. Is it
likely that they contain the same number of genes? A similar number of chromosomes?
6. Which of the following questions would not be answered by using karyotyping?
A) Is the individual genetically female or male?
B) Do any of the chromosomes contain pieces that belong to other chromosomes?
C) Does the individual have an extra chromosome?
D) Do any chromosomes contain point mutations?
7. For each of the following sentences, choose one of the options enclosed in square
brackets to make a correct statement about nucleosomes.
A. Nucleosomes are present in [prokaryotic/eukaryotic] chromosomes, but not in
[prokaryotic/eukaryotic] chromosomes.
B. A nucleosome contains two molecules each of histones [H1 and H2A/H2A and
H2B] as well as of histones H3 and H4.
C. A nucleosome core particle contains a core of histone with DNA wrapped
around it approximately [twice/three times/four times].
D. Nucleosomes are aided in their formation by the high proportion of
[acidic/basic/polar] amino acids in histone proteins.
E. Nucleosome formation compacts the DNA into approximately
[one-third/one-hundredth/one-thousandth] of its original length.
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8. You are studying a newly identified chromatin-remodeling complex, which you call
NICRC. You decide to run an in vitro experiment to characterize the activity of the
purified complex. Your molecular toolbox includes: (1) a 400-base-pair DNA molecule
that has a single recognition site for the restriction endonuclease EcoRI, an enzyme that
cleaves internal sites on double-stranded DNA (dsDNA); (2) purified EcoRI enzyme;
(3) purified DNase I, a DNA endonuclease that will cleave dsDNA at nonspecific sites
if they are exposed; and (4) core octamer histones. You are able to assemble core
nucleosomes on this DNA template and test for NICRC activity. Figure Q5-60A
illustrates the DNA template used and indicates both the location of the EcoRI cleavage
site and the size of the DNA fragments that are produced when it cuts. Figure Q5-60B
illustrates how the DNA molecules in your experiment looked after separation
according to size by using gel electrophoresis. Your experiment had a total of six
samples, each of which was treated according to the legend below the gel. The sizes of
the DNA fragments observed are indicated on the left side of the gel.
Figure Q5-60
A. Explain the results in lanes 1–4 and why it is important to have this information
before you begin to test your remodeling complex.
B. What can you conclude about your purified remodeling complex from the results
in lanes 5 and 6?
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