1. Introduction to Microbiology
Anas Abu-Humaidan M.D. Ph.D.
Lecture 6

2. Top Adenine thymidine
(A-T) pairing; bottom: guanine-cytosine (G-C) pair
100s-1000s kbp
Uracil in ribonucleic acid, or Thymidine in deoxyribonucleic acid
G+C contect

3. Bacterial DNA
Circular chromosomal and extra chromosomal (plasmids, bacteriophages) replicons.
Supercoiled DNA…. No histones, polyamins
Haploid smaller… mutations
The majority of antibiotics that target the 30S subunit inhibit protein synthesis by preventing either the binding of tRNAs to the ribosome or the movement of tRNAs through the ribosome during translocation …. Aminoglycosides and tetracyclines

4. Plasmids
1-200 kbp of circular dsDNA coding for at least one gene. Plasmids can be a Replicon or epitome.
Important in transfer of antibiotic resistance and virulence factors.
Incompatible plasmids (belonging to the same incompatibility group) normally share the same replication or partition mechanisms and can thus not be kept together in a single cel
Fertility F-plasmids, which contain tra genes. They are capable of conjugation and result in the expression of sex pili.
Resistance (R) plasmids, which contain genes that provide resistance against antibiotics or poisons.
Col plasmids, which contain genes that code for bacteriocins, proteins that can kill other bacteria
Virulence plasmids, which turn the bacterium into a pathogen.

5. Transcription and translation
the DNA double helix if read in the3′–5′ direction. Thus, an mRNA is oriented in a 5′–3′direction.
No nulear membrane couple transcription, translation
No poly adenalation (tRNA and rRNA genes mostly at any point) , no introns
Cistrons /genes mostly functional unlike eukaryotes (coding structural proteins genes … mRNA)
Rho-independent termination is controlled by specific sequences in the DNA template strand. As the polymerase nears the end of the gene being transcribed, it encounters a region rich in C–G nucleotides. The mRNA folds back on itself, and the complementary C–G nucleotides bind together. The result is a stable hairpin

6. Transcription and translation
RNA polymerase
The enzyme peptidyltransferase (which is actually the 23S rRNA, ie, a ribozyme) catalyzes the formation of the peptide bond

7. Regulation of transcription
prokaryotic structural genes that encode a related series of metabolic reactions are clustered on operons
prokaryotic structural genes that encode a related series of metabolic reactions are clustered on operons …… polycistronic mrna

8. Regulation of transcription
Repressors are proteins that suppress transcription of a gene in response to an external stimulus, whereas activators are proteins that increase the transcription of a gene in response to an external stimulus.
Inducers are small molecules that either activate or repress transcription depending on the needs of the cell and the availability of substrate
Repressors are proteins that suppress transcription of a gene in response to an external stimulus, whereas activators are proteins that increase the transcription of a gene in response to an external stimulus. Finally, inducers are small molecules that either activate or repress transcription depending on the needs of the cell and the availability of substrate

9. DNA replication

10. DNA replication

11. DNA replication
Because bacterial topoisomerases are essential and unique, they are targets of antibiotics (eg, quinolones).
Because bacterial topoisomerases are essential and unique, they are targets of antibiotics (eg, quinolones).

13. DNA mutations
missense mutation results in a different amino acid being inserted in the protein, but this may be a conservative mutation if the new amino acid has similar properties ….. Null mutations redenreds a gene non/functional
Exogenous triggers include UV irradiation, chemicals or oxidative compounds, acids, organic mutagens, some antibiotics 

14. DNA mutations repair
1. Direct DNA repair is the enzymatic removal of damage, such as pyrimidine dimers and alkylated bases.
2. Excision repair is the removal of a DNA segment containing the damage, followed by synthesis of a new DNA strand.
3. The SOS response is the induction of many genes (≈15) after DNA damage or interruption of DNA replication to promote recombination or error-prone repair.
5. Error-prone repair is the last resort of a bacterial cell before it dies. It is used to fill in gaps with a random sequence when a DNA template is not available for directing an accurate repair.

15. Horizontal Gene transfer (HGT)

16. Gene transfer/ Conjugation

17. Gene transfer/ Transduction

18. Bacteriophages

19. Gene transfer/ Transformation

20. Gene transfer/ Transformation

21. Gene transfer/ Transformation

22. Gene transfer/ Transposition

23. Implications of HGT / antibiotic resistance
The resulting S. aureus plasmid encodes resistance to β-lactams, vancomycin, trimethoprim, and gentamycin/ kanamycin/tobramycin antibiotics

24. DNA cloning

25. Further reading and material:
Murray - Medical Microbiology 8th Edition
Section 4: Bacteriology
Chapter 13: Bacterial metabolisim and genetics
Jawetz, Melnick & Adelberg's Medical Microbiology, 26th edition- Section 1: Fundamentals of Microbiology-
Chapter 7:Microbial genetics
Youtube:
Channel: Judith Leatherman
Video : Bacterial genetics part 1
Video : Bacterial genetics part 2

26. Further reading: Murray - Medical Microbiology 8th Edition
Section 4: Bacteriology
Chapter 13: Bacterial metabolisim and genetics
Jawetz, Melnick & Adelberg's Medical Microbiology, 26th edition- Section 1: Fundamentals of Microbiology-
Chapter 7:Microbial genetics