DNA Structure & History (Ch 6.1/6.
2)
● DNA: Double helix, antiparallel strands (5’→3’ and 3’→5’), deoxyribose sugar, A-T (2
H-bonds), C-G (3 H-bonds).
● RNA: Single-stranded, ribose sugar, U replaces T.
● Nucleotide: Phosphate + sugar (ribose/deoxyribose) + nitrogenous base.
● Purines (2 rings): Adenine, Guanine. Pyrimidines (1 ring): Cytosine, Thymine, Uracil.
● Key Experiments:
○ Griffith: Transformation (R strain + heat-killed S strain → virulent).
○ Avery, McCarty, MacLeod: DNA = transforming factor.
○ Hershey-Chase: Used ³²P (DNA) and ³⁵S (protein) in bacteriophages → DNA is
genetic material.
○ Chargaff: A=T, C=G.
○ Franklin/Wilkins: X-ray diffraction showed helical structure.
○ Watson & Crick: Built DNA model using Chargaff’s rules + Franklin’s data.
DNA Replication (Ch 6.4)
● Semiconservative: Each new DNA molecule = 1 old + 1 new strand (proven by
Meselson-Stahl).
● Enzymes & Steps:
6. Helicase: Unwinds DNA at replication fork.
7. Topoisomerase: Prevents supercoiling.
8. SSBs: Stabilize single strands.
9. Primase: Adds RNA primer.
10.DNA Pol III: Synthesizes DNA (5’→3’).
11.Leading strand: Continuous.
12.Lagging strand: Okazaki fragments (discontinuous).
13.DNA Pol I: Replaces RNA primer with DNA.
14.Ligase: Joins Okazaki fragments.
● Proofreading: DNA Pol III & I correct errors (exonuclease activity).
Transcription & Translation (Ch 7.1-7.3)
Central Dogma: DNA → mRNA → Protein.
● Transcription (nucleus):
○ RNA Pol II binds promoter (TATA box) → synthesizes pre-mRNA.
○ Post-transcriptional modifications:
■ 5’ cap (protection).
■ Poly-A tail (stability).
■ Splicing: Spliceosome removes introns, joins exons.
● Translation (ribosome):
○ tRNA: Brings amino acids; has anticodon matching mRNA codon.
○ Ribosome sites:
■ A site: New tRNA enters.
■ P site: Peptide bond forms.
■ E site: tRNA exits.
○ Start codon (AUG), stop codons (UAA, UAG, UGA).
Gene Regulation (Ch 7.4)
● Lac Operon (inducible):
○ No lactose: Repressor binds operator → no transcription.
○ Lactose present: Binds repressor → transcription.
● Trp Operon (repressible):
○ No tryptophan: Repressor inactive → transcription.
○ Tryptophan present: Corepressor → repressor binds operator → no transcription.
Mutations (Ch 7.5)
● Point Mutations:
○ Silent: No amino acid change.
○ Missense: Amino acid change.
○ Nonsense: Premature stop codon.
● Frameshift: Insertion/deletion (not multiples of 3).
● Causes: Spontaneous (errors), induced (mutagens like UV).
Biotechnology (Ch 8.1-8.2)
● Restriction Enzymes: Cut DNA at specific sequences (e.g., EcoRI → sticky ends).
● Recombinant DNA: Insert gene into plasmid → transform competent cells (CaCl₂ + heat
shock).
● PCR: Amplifies DNA (Denature → Anneal primers → Extend with Taq polymerase).
● Gel Electrophoresis: Separates DNA by size (- → + charge).
DNA Structure & Replication
1. Which of the following is a purine?
a) Cytosine
b) Thymine
c) Adenine
d) Uracil
2. Complementary base pairing in DNA follows which rule?
a) A-G, C-T
b) A-T, C-G
c) A-U, C-G
d) A-C, T-G
3. The enzyme that unwinds DNA during replication is:
a) Helicase
b) Ligase
c) Polymerase
d) Topoisomerase
4. Okazaki fragments are found on the:
a) Leading strand
b) Lagging strand
c) Template strand
d) Coding strand
5. Semiconservative replication means:
a) One old strand and one new strand in each daughter DNA
b) Both strands are completely new
c) DNA is replicated without enzymes
d) Only half of the DNA is copied
6. DNA ligase is responsible for:
a) Unwinding DNA
b) Joining Okazaki fragments
c) Adding RNA primers
d) Proofreading DNA
7. Topoisomerase prevents:
a) DNA breakage
b) Supercoiling
c) Mismatched bases
d) RNA synthesis
8. The replication origin is where:
a) Transcription begins
b) DNA replication starts
c) Translation occurs
d) RNA is spliced
Transcription & Translation
9. The central dogma of molecular biology describes:
a) DNA → RNA → Protein
b) RNA → DNA → Protein
c) Protein → RNA → DNA
d) DNA → Protein → RNA
10.Transcription produces:
a) tRNA
b) mRNA
c) rRNA
d) DNA
11.The TATA box is part of the:
a) Coding strand
b) Promoter
c) Terminator
d) Intron
12.Introns are:
a) Coding sequences
b) Non-coding sequences removed during splicing
c) Part of the final mRNA
d) Found only in prokaryotes
13.The spliceosome is responsible for:
a) Adding the 5’ cap
b) Removing introns
c) Initiating translation
d) Breaking DNA
14.The start codon is:
a) UAA
b) UGA
c) AUG
d) UAG
15.tRNA carries:
a) DNA
b) Amino acids
c) mRNA
d) Ribosomes
16.The anticodon is found on:
a) mRNA
b) tRNA
c) rRNA
d) DNA
Gene Regulation & Mutations
17.The lac operon is activated when:
a) Glucose is present
b) Lactose is present
c) Tryptophan is present
d) Repressor binds DNA
18.A missense mutation results in:
a) A premature stop codon
b) No change in amino acid
c) A different amino acid
d) A frameshift
19.A frameshift mutation is caused by:
a) A single nucleotide substitution
b) Insertion or deletion of nucleotides (not in multiples of 3)
c) A silent mutation
d) A translocation
20.Restriction enzymes cut DNA at:
a) Random sites
b) Specific recognition sites
c) Only blunt ends
d) Only sticky ends
Answer Key
MCQ Answers
1. c
2. b
3. a
4. b
5. a
6. b
7. b
8. b
9. a
10.b
11.b
12.b
13.b
14.c
15.b
16.b
17.b
18.c
19.b
20.b
20 Short Answer Questions
1. Define the role of histones in DNA packaging.
2. What is the function of a plasmid in bacteria?
3. Explain Griffith’s transformation experiment.
4. How does DNA polymerase III differ from DNA polymerase I?
5. Describe the structure of a nucleosome.
6. What is the Hayflick limit?
7. How does telomerase prevent cell senescence?
8. Explain the difference between exons and introns.
9. What is the function of the 5’ cap and poly-A tail in mRNA?
10.Describe how the lac operon is regulated.
11.What is the difference between a missense and nonsense mutation?
12.How does PCR amplify DNA?
13.What is the purpose of gel electrophoresis?
14.Explain how a restriction enzyme creates sticky ends.
15.What is recombinant DNA?
16.How does a repressor protein regulate the trp operon?
17.What is the reading frame in translation?
18.How does aminoacylation charge a tRNA?
19.What is the role of snRNPs in splicing?
20.Why are Okazaki fragments necessary in DNA replication?
Short Answer Key
1. Histones help package DNA into chromatin.
2. Plasmids are small, circular DNA molecules used for gene transfer in bacteria.
3. Griffith’s experiment showed that a "transforming factor" (DNA) could change harmless
bacteria into pathogenic ones.
4. DNA Pol III synthesizes new DNA, while DNA Pol I removes RNA primers and fills gaps.
5. Nucleosome = DNA wrapped around 8 histone proteins.
6. Hayflick limit = Maximum number of divisions a cell can undergo before senescence.
7. Telomerase extends telomeres, preventing chromosomal shortening.
8. Exons are coding regions; introns are non-coding and spliced out.
9. 5’ cap protects mRNA; poly-A tail stabilizes it.
10.Lac operon: Lactose inactivates repressor → transcription.
11.Missense = amino acid change; nonsense = stop codon.
12.PCR cycles denaturation, annealing, and extension to amplify DNA.
13.Gel electrophoresis separates DNA by size.
14.Restriction enzymes cut DNA unevenly, leaving overhangs (sticky ends).
15.Recombinant DNA = DNA combined from different sources.
16.Trp repressor binds operator when tryptophan is present, blocking transcription.
17.Reading frame = correct triplet grouping for translation.
18.Aminoacylation attaches an amino acid to tRNA.
19.snRNPs help splice introns from pre-mRNA.
20.Okazaki fragments allow discontinuous synthesis on the lagging strand.