GENETICS QUESTIONS DNA & REPLICATION

1. 2. 3. What are the chemical components of DNA? Explain the term DNA replication Define the following terms, used to describe the structure of DNA: a. phosphodiester bond b. complementary base pairing c. antiparallel d. hybridization What is the role of each enzyme involved in DNA replication? The nucleotide sequence of a region of one DNA strand of a DNA double helix is: 5'-GACTTACGACTAG-3' What is the sequence of the complementary strand? The strands of a DNA double helix can be separated ("denatured") by heating. If you raised the temperature of a solution containing the following 3 DNA fragments in what order will they denature? Explain your answer. (a) 5'-AGCCGATCCG-3' 3'-TCGGCTAGGC-5' (b) 5'-ATTTAAGATTCA-3' 3'-TAAATTCTAAGT-5' (c) 5'-GCGCACGCTGG-3' 3'-CGCCTGCGACC-5' About 21% of the bases in rat DNA are guanine. What are the percentages of the other bases? Explain your answer.

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Meselson and Stahl carried out the following experiment that demonstrated that DNA replication is a semi-conservative process: The DNA of Escherichia coli was "labelled" by growing the bacteria in a medium in which the nitrogen source was 15N so that both DNA strands were "heavy". The bacteria were then transferred to a medium containing 14N so that all subsequent DNA chains would be "light". After 3 generations (i.e. 3 rounds of replication) the DNA was analysed by density gradient centrifugation. Which of the following density gradient patterns would be observed? Explain your answer. (N.B. the area under the curve is a measure of the amount of double-stranded DNA with a particular composition of 15N- and 14N-labelled chains)

GENETICS QUESTIONS DNA & REPLICATION ANSWERS

1. What are the chemical components of DNA? Nitrogenous bases (adenine, guanine, cytosine & thymine), deoxyribose sugar , phosphate 2. Explain the term DNA replication synthesis of new DNA strands using a DNA template 3. Define the following terms, used to describe the structure or properties of DNA: a. phosphodiester bond a covalent bond in DNA and RNA between twosugars and a phosphate b. complementary base pairing the hydrogen bonding between a particular purine and a particular pyrimidine in double-stranded nucleic acid molecules (DNADNA, DNA-RNA or RNA-RNA) c. antiparallel the orientation of the two polynucleotide chains of a double helix: one runs in a 5 to 3 direction and the other in a 3- to 5 direction d. hybridisation the assocation of two complementary single-stranded polynucleotides to form a double helix 4. What is the role of each enzyme involved in DNA replication? 5. The nucleotide sequence of a region of one DNA strand of a DNA double helix is: 5'-GACTTACGACTAG-3' What is the sequence of the complementary strand? 3-CTGAATGCTGATC-5 6. The strands of a DNA double helix can be separated ("denatured") by heating. If you raised the temperature of a solution containing the following 3 DNA fragments in what order will they denature? Explain your answer. (a) 5'-AGCCGATCCG-3' 3'-TCGGCTAGGC-5' (b) 5'-ATTTAAGATTCA-3' 3'-TAAATTCTAAGT-5' (c) 5'-GCGCACGCTGG-3' 3'-CGCCTGCGACC-5' Fragment (b) denatures first then fragment (a) and finally fragment(c) The heat stability of the double-stranded fragments is determined by the number of hydrogen bonds between the two chains of the fragments [(a) has 27 H-bonds, (b) has 26 H-bonds, (c) has 31 H-bonds]. There are 2 H-bonds in an A-T pair and 3 Hbonds in a G-C pair. 7. About 21% of the bases in rat DNA are guanine. What are the percentages of the other bases? Explain your answer. In double-stranded DNA the adenine content equals that of the thymine content and the guanine content equals that of the cytosine content because of the rules of base pairing. Therefore rat DNA must also contain 21% cytosine, 29% adenine and 29% thymine. You can check your result because the sum of the purines (A+G) must equal that of the sum of the pyrimidines (T+C).

GENETICS QUESTIONS DNA & REPLICATION

8. Meselson and Stahl carried out the following experiment that demonstrated that DNA replication is a semi-conservative process: The DNA of Escherichia coli was "labelled" by growing the bacteria in a medium in which the nitrogen source was 15N so that both DNA strands were "heavy". The bacteria were then transferred to a medium containing 14N so that all subsequent DNA chains would be "light". After 3 generations (i.e. 3 rounds of replication) the DNA was analysed by density gradient centrifugation. Which of the following density gradient patterns would be observed? Explain your answer. (N.B. the area under the curve is a measure of the amount of double-stranded DNA with a particular composition of 15N- and 14N-labelled chains)

Pattern (c ) would be observed. After 1 generation all the DNA chains will be identical and contain one heavy DNA chain from the original 15N-labelled double-stranded DNA and one new light DNA chain (i.e. all the DNA will be a hybrid of 15N and 14N chains). After 2 generations half the DNA will be exclusively 14N-labelled (the 14N-labelled chains synthesised after 1 generation act as template for the synthesis of a complementary 14Nlabelled chain), the original 15N-labelled chains will again act as templates for the synthesis of complementary 14Nlabelled chains (i.e. the DNA will be a 50:50 mixture of 15N/14N DNA and 14N/14N DNA). After 3 generations once again the 15N-labelled DNA chains will form 15N/14N hybrids and the remaining DNA chains will only comprise light chains. 25% of the DNA will now be 15N/14N hybrids and 75% of the DNA 14N/14N (i.e. the DNA will be a mixture of 15N/14N DNA and 14N/14N DNA in the ratio 1:3).