Micro Test 2 condensed – Flashcards
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DNA to mRNA protein synthesis biosynthesis |
Initiation requirements |
promoter RNA (pol)ymerase Sigma (σ) Factor |
RNA Holoenzyme |
RNA pol + σ factor
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RNA pol |
synthesizes RNA 5 protein subunits |
Inititation |
1. RNA Holoenzyme binds to DNA (forms Closed Complex) 2. DNA is unwound (Start of Open confirmation) |
Once Elongation starts |
σ factor falls off |
Initiation of Translation |
1. Ribosome binds to mRNA at Shine-Delgarno sequence 2. AUG is first codon
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Shine Delgarno sequence |
Ribosomal binding site of for inititation of translation |
AUG codon is |
methionine |
Initiation Complex is composed of |
30S + 50S subunits + mRNA + fMet-tRNA |
Elongation Peptide formation 3 steps |
1. Aminoacyl-tRNA arrives at A site 2. Transpeptidation reaction occurs between P and A sites 3. Translocation – Empty tRNA exits from E site – tRNA with growing chain moves from A to P – New tRNA comes into the A site
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Ribosome has 3 binding sites |
1. E site (exit site) 2. P site (peptidyl, or donor site) 3. A site (aminoacyl, or acceptor)
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Some proteins modified after synthesis by adding |
+lipids= lipoproteins; gram - +sugars= glycoproteins; gram + |
Operons |
two or more genes transcribed from a single promoter |
Regulatory Regions |
• Regulatory regions are upstream of genes; one regulatory region can control grouped genes
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3 levels of control |
Transcriptional- change in gene expression levels by altering transcription rates. Translational - control of the levels of protein synthesized from its mRNA. Protein- how much the mRNA is translated into proteins. |
Transcriptional Control example |
Diauxic Growth of E. coli-
transition occurs; glucose genes off, lactose genes on
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Positive Control |
when a regulatory protein promotes transcription initiation; activator protein binds upstream and promotes RNA Holoenzyme binding
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Negative Control |
when a regulatory protein inhibits transcription initiation; repressor protein binds to operator to prevent transcription
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Types of Genetic Variability |
Mutation: induced/spontaneous Recombination: leads to newcombintations of genes on a chromosome |
Silent Mutation |
point mutation alteration of base pair resulting in SAME amino acid |
Prion
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infectious proteins that lack nucleic acids mad cow |
Missense Mutation |
point mutation alteration of base pair resulting in DIFFERENT amino acid; |
Frame Shift Mutation |
addition/removal of a base; results in a shift in codon reading and alters protein function |
Mutation phenotypes |
Auxotroph- nutritional mutant that requires an additional growth factor Prototroph- parent of an auxotroph
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Motility mutation |
loss of flagella |
Temperature sensitive mutation |
mutant with a protein that becomes denatured at a certain temperature |
resitant mutation |
antibiotics, heavy metals, phage |
Ames Test |
Fast, easy to idenitify mutagens; an increase in reversion rate = potential mutagen/carcinoge |
Conjugation |
bacterial sex plasmid DNA carries gene for sex pilus; F+ cell-cell contact required |
Bacterial Growth Conditions |
– Temperature – Nutrients – Water (osmotic balance) – Appropriate atmosphere – pH |
Bacteriostatic |
stops bacterial growth -static/-stasis = standing still |
Microbial Death |
D value; decimal reduction time time require to kill 90% of microbes or spores |
anti microbial targets |
ribosomes DNA Enzymes Cell wall Membranes |
what affects D-Value |
intensity/concentration- lowers D-value Microbial load- number of microbes; D-value same rate, increased time to do so Population Compostition- mixture of microbes Temperature- increased temperature, more active Environment- type of material contaminated |
high temperature physical agents |
autoclave- moist heat, under pressure. More quantity, more time Boiling- moist heat, not under pressure pasteurization- sub-boiling temps; high temp short time, ultra high temp, Hot air incinteration |
Dessication |
freeze dry |
radiation |
physical DNA damaged ionzing- gamma rays/xrays non-ionizing- UV light |
Filtration |
physical seperates bacteria from liquid HEPA filtration for air ultra filtration- filters out lipopolysaccharide endotoxins from dead gram - |
phenolic disinfectant |
chemical Bactericidal or bacteriostatic depending on concentration higher concentration = bacterialcidal – Increases membrane permeability – Denatures protein
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Halogens as disinfectants |
Chemical Oxidizing agent, inactivates key proteins
Iodine – very effective germicide Chlorine
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Heavy Metals as disinfectants |
chemical
• Antibacterial and effective against algae and fungi • Mode of action – Inactivate enzymes - react with –SH group toxic
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Detergents as disinfectants |
chemical Surface active compounds (“surfactants”) – Damage cell membranes – Denature protein
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Quaternary Ammonium Compounds |
chemical disinfecting, sanitizing, and antisepsis |
Antibiotics |
compounds (natural or synthetic) that inhibit specific metabolic processes in cells Paul Ehrlich- syphillis antibiotic Alexander Fleming- penecillin
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antibiotic targets
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• Cell wall synthesis • Membrane function • Protein synthesis • Metabolite antagonism
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inhibitors of cell wall synthesis |
Active against Gram positives – Natural or semi-synthetic – Allergies and pathogen resistance ex PENECILLIN |
Inhibitors of DNA Function |
Quinolones (nalidixic acid, ciprofloxacin) – Inhibit DNA gyrase; Very broad spectrum Mitomycin – Cross-links with DNA; Induces endonuclease activity
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Antibiotic Metabolite Analogs |
Enzyme inhibitors ex. Sulfonamide |
Saprophytic fungi |
decompose organic matter |
Fungi cause Disease in |
• Humans and animals • Plant pathogens • Pests
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marine phytoplankton are responsible for |
CO2 fixation and O2 evolution on Earth
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Lichens
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a unique symbiotic life form consisting of fungi and algae (actually cyanobacteria)
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Blue-Green Algae are |
prokaryotic thus call Cyanobacteria |
Protozoa |
Protozoology: study of protozoa Eukaryotic, unicellular animals Motile by cilia, flagella, or amoeboid motility
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Virus Characteristics |
Require host machinery to multiply 1 nucleic acid- DNA or RNA acellular capsid- some have lipid envelope Nucelocapsid Virion- complete individual virus |
Virus size/shape |
10nm - 300nm helical- tube shape polyhedral- multi-sided
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lytic bacteriophage infection process |
• Attachment: adhesion to target site • Entry: Virus injects nucleic acid into host • Replication: early viral proteins shut down host metabolism, then replicates • Maturation/assembly: Capsids/nucleic acids; sometimes host DNA picked up • Release: Virions are released; Cell lysis; budding |
lysogenic phase process |
attachment/entry- same; except DNA becomes circular Repression- phage represses lytic cycle Integration- phage inserts into host DNA; prophage attainment- prophage replicates with chromosome; repressor protein blocks lytic Entry- to lytic cycle; prophage expressed
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Plant and animal viruses |
contain envelopes with Glycoprotein spikes 'naked' virion = non-enveloped
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oncogenic viruses |
Incorporation of viral DNA into host DNA may interrupt cell function |
Treatments for Viruses |
Immune system- immunity from current/previous infection. Vaccination: exposure to virus (inactivated or portion of virus) Fever: increased temp inactivates some viral proteins Interferon: antiviral produced by the body, blocks viral protein synthesis |