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Biology 30 · Molecular genetics

DNA up close

The genetics unit showed how traits are inherited. This one zooms in: how DNA copies itself, how its code becomes a protein, what happens when a letter changes, and how scientists cut, copy and sort it.

DNA structure and replication

The idea: DNA is two strands of nucleotides held together by paired bases. To copy it, the strands separate and each one is used as a template.

Each nucleotide has a phosphate, a deoxyribose sugar and a base. Sugar and phosphate form the backbone; the bases pair across the middle: A with T and C with G.

Chargaff's rule

Because the bases pair, A = T and C = G in any DNA sample. If 30% is adenine, then thymine is also 30%, leaving 40% split evenly: 20% guanine and 20% cytosine.

Replication, step by step

  1. Helicase unwinds the helix and separates the strands.
  2. DNA polymerase adds matching nucleotides to each template strand.
  3. It can only build in one direction, so one strand (leading) is made continuously and the other (lagging) is made in short Okazaki fragments.
  4. DNA ligase joins the fragments.

Every new molecule keeps one original strand and gets one new one — that is why replication is called semi-conservative.

Protein synthesis

The idea: transcription copies a gene into mRNA in the nucleus; translation reads the mRNA three bases at a time at a ribosome to build a protein.

TranscriptionTranslation
WhereNucleusRibosome
MakesmRNA from a DNA templateA chain of amino acids from mRNA
Key playersRNA polymerasemRNA codons, tRNA anticodons, ribosomes
  • mRNA processing: in eukaryotes, non-coding introns are cut out and the exons are joined before the mRNA leaves the nucleus.
  • Codons: each three-base mRNA codon means one amino acid. AUG is the start codon; UAA, UAG and UGA are stop codons.
  • tRNA: carries one amino acid and has an anticodon that pairs with the codon. The anticodon for AUG is UAC.
  • Redundancy: 64 codons but only 20 amino acids, so many amino acids have several codons.

Watch out: writing T in any RNA sequence. RNA uses U instead of T, in both codons and anticodons.

Mutations

The idea: how much a mutation matters depends on what it does to the codons — change nothing, change one amino acid, stop the protein, or scramble everything after it.

TypeWhat happensExample
SilentA substitution, but the codon still means the same amino acidGAA → GAG (both glutamic acid)
MissenseA substitution changes one amino acidSickle cell anemia
NonsenseA substitution creates a stop codon, cutting the protein shortUAC → UAA
FrameshiftAn insertion or deletion shifts the reading frame, changing every codon after itUsually a non-working protein

Mutations can happen by chance during replication or be caused by mutagens such as UV light, X-rays and some chemicals. They are also the original source of every new allele — without them, there would be no variation for evolution to act on.

Biotechnology

The idea: four tools do most of the work — enzymes that cut DNA, enzymes that join it, a way to copy it, and a way to sort it by size.

ToolWhat it does
Restriction enzymesCut DNA at specific sequences, often leaving "sticky ends"
DNA ligaseJoins pieces of DNA together
PCRMakes millions of copies of a DNA segment by repeated heating and cooling
Gel electrophoresisSeparates DNA fragments by size
laddersample 1sample 2 + larger smaller
DNA is negatively charged, so it moves toward the positive end. Small fragments slip through the gel faster and end up farthest from the wells. The ladder of known sizes lets you measure the samples.

Making human insulin in bacteria

  1. Cut the human insulin gene and a bacterial plasmid with the same restriction enzyme, so their sticky ends match.
  2. Join them with DNA ligase, making recombinant DNA.
  3. Put the plasmid into bacteria. As they multiply, they read the human gene and make insulin.

The same toolkit underlies DNA fingerprinting in forensics, genetically modified crops, gene therapy research and newer gene-editing tools like CRISPR — each of which raises its own ethical questions.

Watch out: PCR copies DNA; gel electrophoresis sorts it. Questions often put one in place of the other.

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