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Microbial Genetics
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Microbial Genetics

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  1. Microbial Genetics

  2. Genetics – the study of heredity • transmission of biological traits from parent to offspring • expression & variation of those traits • structure & function of genetic material • how this material changes

  3. Levels of genetic study

  4. Levels of structure & function of the genome • genome – sum total of genetic material of an organism (chromosomes + mitochondria/chloroplasts and/or plasmids) • genome of cells – DNA • genome of viruses – DNA or RNA • chromosome – length of DNA containing genes • gene-fundamental unit of heredity responsible for a given trait • site on the chromosome that provides information for a certain cell function • segment of DNA that contains the necessary code to make a protein or RNA molecule

  5. Genomes vary in size • smallest virus – 4-5 genes • E. coli – single chromosome containing 4,288 genes; 1 mm; 1,000X longer than cell • Human cell – 46 chromosomes containing 31,000 genes; 6 feet; 180,000X longer than cell

  6. Nucleic acids are made of nucleotides similar to how proteins are made of amino acids • each nucleotide consists of 3 parts • a 5 carbon sugar (deoxyribose or ribose) • a phosphate group • a nitrogenous base (adenine, thymine, cytosine, guanine, and uracil)

  7. DNA structure • 2 strands twisted into a helix • sugar -phosphate backbone • nitrogenous bases form steps in ladder • constancy of base pairing • A binds to T with 2 hydrogen bonds • G binds to C with 3 hydrogen bonds • antiparallel strands 3’to 5’ and 5’to 3’ • each strand provides a template for the exact copying of a new strand • order of bases constitutes the DNA code

  8. Significance of DNA structure • Maintenance of code during reproduction. Constancy of base pairing guarantees that the code will be retained. • Providing variety. Order of bases responsible for unique qualities of each organism.

  9. DNA replication is semiconservative because each chromosome ends up with one new strand of DNA and one old strand.

  10. Semi-conservative replication of DNA

  11. DNA replication • Begins at an origin of replication • Helicase unwinds and unzips the DNA double helix • An RNA primer is synthesized • DNA polymerase III adds nucleotides in a 5’ to 3’ direction • Leading strand – synthesized by DNA polymerase continuously in 5’ to 3’ direction • Lagging strand – synthesized 5’ to 3’ in short segments; overall direction is 3’ to 5’ • DNA is read in the 3’ to 5’ direction; Nucleotides are added in the 5’ to 3’ direction.

  12. Bacterial replicon

  13. Flow of genetic information

  14. What are the products that genes encode? • RNAs and proteins • How are genes expressed? • transcription and translation

  15. Gene expression • Transcription – DNA is used to synthesize RNA • RNA polymerase is the enzyme responsible • Translation –making a protein using the information provided by messenger RNA • occurs on ribosomes

  16. Genotype - genes encoding all the potential characteristics of an individual • Phenotype -actual expressed genes of an individual (its collection of proteins)

  17. DNA-protein relationship • Each triplet of nucleotides (codon) specifies a particular amino acid. • A protein’s primary structure determines its shape & function. • Proteins determine phenotype. Living things are what their proteins make them. • DNA is mainly a blueprint that tells the cell which kinds of proteins to make and how to make them.

  18. DNA-protein relationship

  19. 3 types of RNA • messenger RNA (mRNA) • transfer RNA (tRNA) • ribosomal RNA (rRNA)

  20. DNA Transcription RNA polymerase RNA Translation ribosomes PROTEINS

  21. Transcription • RNA polymerase binds to promoter region upstream of the gene • RNA polymerase adds nucleotides complementary to the template strand of a segment of DNA in the 5’ to 3’ direction • Uracil is placed as adenine’s complement • At termination, RNA polymerase recognizes signals and releases the transcript • 100-1,200 bases long

  22. Transcription

  23. Translation • Ribosomes assemble on the 5’ end of a mRNA transcript • Ribosome scans the mRNA until it reaches the start codon, usually AUG • A tRNA molecule with the complementary anticodon and methionine amino acid enters the P site of the ribosome & binds to the mRNA

  24. Translation

  25. Using this chart, you can determine which amino acid the codon “codes” for! Which amino acid is encoded in the codon CAC?

  26. Find the second letter of the codon CAC Find the first letter of the codon CAC Find the third letter of the codon CAC

  27. CAC codes for the amino acid histidine (his).

  28. What does the mRNA codon UAC code for? Tyr or tyrosine

  29. Interpreting the DNA code

  30. Translation elongation • A second tRNA with the complementary anticodon fills the A site • A peptide bond is formed • The first tRNA is released and the ribosome slides down to the next codon. • Another tRNA fills the A site & a peptide bond is formed. • This process continues until a stop codon is encountered.

  31. Translation termination • Termination codons – UAA, UAG, and UGA – are codons for which there is no corresponding tRNA. • When this codon is reached, the ribosome falls off and the last tRNA is removed from the polypeptide.

  32. Polyribosomal complex

  33. Eucaryotic transcription & translation differs from procaryotic • Do not occur simultaneously. Transcription occurs in the nucleus and translation occurs in the cytoplasm. • Eucaryotic start codon is AUG, but it does not use formyl-methionine. • Eucaryotic mRNA encodes a single protein, unlike bacterial mRNA which encodes many. • Eucaryotic DNA contains introns – intervening sequences of noncoding DNA- which have to be spliced out of the final mRNA transcript.

  34. Split gene of eucaryotes

  35. Multiplication of dsDNA viruses

  36. Multiplication of +ssRNA

  37. Regulation of protein synthesis & metabolism

  38. Operons • a coordinated set of genes, all of which are regulated as a single unit. • 2 types • inducible – operon is turned ON by substrate: catabolic operons- enzymes needed to metabolize a nutrient are produced when needed • repressible – genes in a series are turned OFF by the product synthesized; anabolic operon –enzymes used to synthesize an amino acid stop being produced when they are not needed