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Chapter 27A: Bacteria and Archaea 1. Extracellular Prokaryotic Structures 2. Intracellular Prokaryotic Structures 3. Genetic Diversity Prokaryotes 1. Extracellular Prokaryotic Structures Prokaryotic Cell Shape cocci bacilli spirilla • spherical prokaryotes are referred to as cocci (singular = coccus) • rod-shaped prokaryotes are referred to as bacilli (singular = bacillus) Spherical 3 µm 1 µm 1 µm • rod-shaped cells can also be slightly curved (vibrio) or spiral (spirillum) or highly coiled (spirochete) Rod-shaped Spiral vibrio spirochete Bacterial Cell Wall Structure (a) Gram-positive bacteria (b) Gram-negative bacteria Carbohydrate portion of lipopolysaccharide Cell wall Peptidoglycan layer Plasma membrane Peptidoglycan traps crystal violet, which masks the safranin dye. Cell wall Outer membrane Peptidoglycan layer Plasma membrane Crystal violet is easily rinsed away, revealing the red safranin dye. Gram Staining A Gram stain is a very common stain to distinguish 2 bacterial types: Gram negative 1 2 3* 4 • does NOT retain crystal violet, only the safranin Gram-negative bacteria Gram positive Gram-positive bacteria • retains crystal violet due to thick peptidoglycan layer 10 µm * key step A Gram stain of 2 distinct Bacterial Species Capsules A polysaccharide or protein layer called a capsule covers many prokaryotes. • mediates adhesion and the formation of biofilms Bacterial cell wall • protects the cell from dessication (drying out) and phagocytosis (being consumed by cells of the immune system) Bacterial capsule Tonsil cell 200 nm Bacterial Flagella • consist of a basal body, hook & filament Flagellum Filament Hook Motor Cell wall Plasma membrane Rod Peptidoglycan layer 20 nm • basal body anchors flagellum in the membrane and cell wall, and rotates the hook & filament to propel bacterium Fimbriae Some prokaryotes have fimbriae (aka “pili”), which allow them to stick to their substrate or other individuals in a colony. Fimbriae 1 µm Sex Pilus Sex pili are longer than fimbriae and allow prokaryotes to exchange DNA through a process called conjugation. Sex pilus 1 µm 2. Intracellular Prokaryotic Structures Endospores Some Gram-positive bacteria form metabolically inactive endospores which can remain viable in harsh conditions for centuries. Endospore • inactive, dormant cells enclosed in a highly resistant spore coat Coat • remain dormant until conditions improve 0.3 µm • very resistant to heating, freezing, dessication and damaging radiation (e.g. UV) Infoldings of the Cell Membrane Some prokaryotes have highly folded membranes to increase the surface area for processes such as cellular respiration and photosynthesis. 1 µm 0.2 µm • such membrane infoldings are not considered to be true organelles such as those found in eukaryotes Respiratory membrane Thylakoid membranes (a) Aerobic prokaryote (b) Photosynthetic prokaryote Prokaryotic Chromosome Prokaryotes typically have 1 circular DNA molecule that is the chromosome. • the prokaryotic chromosome is located in a region of the cell called the nucleoid Plasmids Some bacteria have 1 or more small, extrachromosomal, non-essential circular DNA molecules called plasmids. Chromosome Plasmids generally contain genes that confer some sort of advantage for survival and reproduction: Plasmids • protection from toxic substances (antibiotic resistance) 1 µm • toxins to kill competitors, enhance disease • gene transfer by conjugation 3. Genetic Diversity in Prokaryotes Sources of Genetic Diversity in Prokaryotes 3 general factors contribute to prokaryotic diversity: MUTATION • changes in DNA sequences RAPID REPRODUCTION • some prokaryotes can reproduce every 20 minutes HORIZONTAL GENE TRANSFER • transfer of DNA from one cell to another Horizontal vs Vertical Gene Transfer Vertical • transfer to the next generation Horizontal (or lateral) • transfer within the same generation Mechanisms of Prokaryotic Gene Transfer Bacteria can acquire DNA (i.e., new genes) in 3 basic ways: 1) Transformation – the uptake and retention of external DNA molecules 2) Conjugation – direct transfer of DNA from one bacterium to another • “regular” conjugation • HFR conjugation 3) Transduction – the transfer of DNA between bacteria by a virus Transformation Under the right conditions, bacteria can “take in” external DNA fragments (or plasmids) by transformation. • DNA binding proteins transfer external DNA across cell wall and membrane • recombination with the chromosomal DNA can then occur Bacterial Conjugation F plasmid Bacterial chromosome F+ cell F+ cell (donor) Mating bridge F− cell (recipient) 1 One strand of F+ cell plasmid DNA breaks at arrowhead. Bacterial chromosome 2 Broken strand peels off and enters F− cell. F+ cell 3 Donor and recipient cells synthesize complementary DNA strands. Conjugation and transfer of an F plasmid 4 Recipient cell is now a recombinant F+ cell. Hfr Conjugation Hfr cell (donor) A+ A+ A+ A− A+ F factor A− F− cell (recipient) 1 An Hfr cell forms a mating bridge with an F− cell. A− 2 A single strand of the F factor breaks and begins to move through the bridge. A+ A− 3 Crossing over can result in exchange of homologous genes. A+ Recombinant F− bacterium 4 Enzymes degrade and DNA not incorporated. Recipient cell is now a recombinant F− cell. Conjugation and transfer of part of an Hfr bacterial chromosome, resulting in recombination Phage DNA 1 Phage infects bacterial donor cell with A+ and B+ alleles. A+ B+ Donor cell 2 Phage DNA is replicated and proteins synthesized. A+ B+ 3 Fragment of DNA with A+ allele is packaged within a phage capsid. A+ 4 Phage with A+ allele infects bacterial recipient cell. 5 Incorporation of phage DNA creates recombinant cell with genotype A+ B−. Transduction Crossing over A+ A− B− Recombinant cell Recipient cell A+ B− A bacteriophage virus can transfer DNA fragments from one host cell to another followed by recombination: • requires a virus to be packaged with bacterial DNA “by mistake” • infection of another cell by such a virus facilitates the gene transfer followed by recombination