🧬 Chapter 8: Microbial Genetics
1. Genes Code for Proteins
Genetics – study of heredity; how genes carry and express information.
Genome – all genetic material (chromosomes + plasmids).
Gene – segment of DNA that codes for a functional product (protein or RNA).
Central Dogma – DNA → RNA → Protein.
Genotype – genetic makeup (potential).
Phenotype – expressed traits (actual functions).
DNA expression → protein synthesis → cellular function.
2. DNA Replication
Purpose: Produces two identical DNA molecules before cell division (vertical gene
transfer).
Process:
o Unwinding: Helicase separates strands; topoisomerase/gyrase relieves
supercoiling.
o Complementary base pairing: A–T, G–C.
o Synthesis: DNA polymerase adds nucleotides to the 3′ end only.
o Leading strand: Synthesized continuously.
o Lagging strand: Made discontinuously (Okazaki fragments) joined by DNA
ligase.
o Semiconservative replication: Each new molecule has one parental and one new
strand.
Energy: Comes from nucleoside triphosphates (ATP, GTP, CTP, TTP).
Accuracy: Proofreading by DNA polymerase keeps errors ~1 in 10 billion bases.
Key Enzymes (Table 8.1):
o Helicase – unwinds DNA
o DNA Polymerase – synthesizes and proofreads
o DNA Ligase – joins fragments
o Gyrase/Topoisomerase – relaxes supercoiling
o Primase – makes RNA primers
o Methylase, Endonuclease, Exonuclease, Photolyase – repair roles.
3. Transcription
Definition: Synthesis of complementary RNA from DNA template.
Steps:
o Initiation: RNA polymerase binds to promoter.
o Elongation: RNA polymerase adds RNA nucleotides (A–U, G–C pairing).
o Termination: Occurs at terminator sequence.
Products:
o mRNA – carries genetic code to ribosome.
o tRNA – transfers amino acids.
o rRNA – part of ribosome.
In Prokaryotes: Occurs in cytoplasm; transcription and translation are coupled.
In Eukaryotes: Occurs in nucleus; RNA processing removes introns using snRNPs,
producing mature mRNA.
4. Translation
Definition: Decoding mRNA into a chain of amino acids (protein synthesis).
Key Terms:
o Codon: 3-base sequence on mRNA specifying an amino acid.
o Start codon: AUG (methionine).
o Stop codons: UAA, UAG, UGA.
o Anticodon: 3-base tRNA sequence complementary to codon.
Process:
o Ribosome assembles on mRNA at start codon.
o tRNAs bring amino acids; peptide bonds form between them.
o Ribosome moves 5′→3′, elongating polypeptide.
o Stops at stop codon → protein released.
Degeneracy: Multiple codons code for same amino acid → reduces effects of mutations.
Prokaryotes: Translation can begin before transcription is complete.
5. Operons and Gene Regulation
Operon: A promoter, operator, and structural genes transcribed together.
Types:
o Inducible Operon (e.g., lac operon):
Normally off; repressor binds operator.
Presence of inducer (allolactose) → repressor inactivated → transcription
on.
o Repressible Operon (e.g., trp operon):
Normally on; excess product (tryptophan) acts as corepressor, activating
repressor → transcription off.
Positive Regulation:
o CAP + cAMP enhance RNA polymerase binding.
o Catabolite Repression: High glucose = low cAMP → lac operon off.
Epigenetic Control: DNA methylation turns genes off (heritable but reversible).
Post-transcriptional Control:
o Riboswitches and miRNA/siRNA regulate mRNA stability and translation.
6. Mutations
Definition: Permanent change in DNA sequence.
Types:
o Base substitution (point mutation): one base replaced.
Missense mutation: changes amino acid (e.g., sickle cell anemia).
Nonsense mutation: introduces stop codon → truncated protein.
Silent mutation: no effect due to genetic code degeneracy.
o Frameshift mutation: insertion or deletion shifts reading frame → nonfunctional
protein.
Spontaneous vs Induced:
o Spontaneous – replication errors.
o Induced – caused by mutagens (chemical or radiation).
7. Plasmids and Recombination
Plasmids: Small circular DNA molecules separate from the chromosome.
o Carry genes for antibiotic resistance, toxins, or conjugation.
Gene Transfer Mechanisms:
o Transformation: Uptake of DNA fragments from environment.
o Conjugation: Direct transfer via pili between bacterial cells.
o Transduction: Transfer by bacteriophage (virus).
Recombination: Integration of foreign DNA into host chromosome → genetic diversity.
Applications: Basis of recombinant DNA technology and genetic engineering.
8. Applications of DNA Biochemistry
Chemotherapy: Target bacterial replication enzymes (e.g., DNA gyrase inhibitors).
Molecular Diagnostics: Detect mutations, resistance genes, or cancer markers (e.g.,
stool DNA test for colorectal cancer).
HIV Treatment: Inhibitors targeting reverse transcription.
Recombinant DNA: Production of human insulin, vaccines, and gene therapy.