Micro Test 2 condensed – Flashcards

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Transcription is
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DNA to mRNA

protein synthesis

biosynthesis

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Initiation requirements
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promoter

RNA (pol)ymerase

Sigma (σ) Factor

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RNA Holoenzyme
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RNA pol + σ factor

 

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RNA pol
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synthesizes RNA

5 protein subunits

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Inititation
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1. RNA Holoenzyme binds to DNA (forms Closed Complex)

2. DNA is unwound

(Start of Open confirmation)

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Once Elongation starts
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σ factor falls off
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Initiation of Translation
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1. Ribosome binds to mRNA at Shine-Delgarno sequence

2. AUG is first codon

 

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Shine Delgarno sequence
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Ribosomal binding site of for inititation of translation
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AUG codon is
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methionine
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Initiation Complex is composed of
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30S + 50S subunits + mRNA + fMet-tRNA
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Elongation Peptide formation 3 steps
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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
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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
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+lipids= lipoproteins; gram -

+sugars= glycoproteins; gram +

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Operons
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two or more genes transcribed from 

a single promoter

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Regulatory Regions
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• Regulatory regions are upstream of 

genes; one regulatory region can control grouped genes

 

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3 levels of control
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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.

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Transcriptional Control example
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Diauxic Growth of E. coli-

 

transition occurs; glucose genes off, lactose genes on

 

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Positive Control
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when a regulatory protein promotes transcription initiation; activator protein binds upstream and promotes RNA Holoenzyme binding

 

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Negative Control
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when a regulatory protein inhibits transcription initiation; repressor protein binds to operator to prevent transcription

 

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Types of Genetic Variability
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Mutation: induced/spontaneous

Recombination: leads to newcombintations of genes on a chromosome

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Silent Mutation
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point mutation

alteration of base pair resulting in SAME amino acid

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Prion

 

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infectious proteins that lack nucleic 

acids

mad cow

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Missense Mutation
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point mutation

alteration of base pair resulting in DIFFERENT amino acid;

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Frame Shift Mutation
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addition/removal of a base; results in a shift in codon reading and alters protein function
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Mutation phenotypes
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Auxotroph- nutritional mutant that requires an additional growth factor

Prototroph- parent of an auxotroph

 

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Motility mutation
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loss of flagella
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Temperature sensitive mutation
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mutant with a protein that becomes denatured at a certain temperature
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resitant mutation
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antibiotics, heavy metals, phage
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Ames Test
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Fast, easy to idenitify mutagens; an increase in reversion rate = potential 

mutagen/carcinoge

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Conjugation
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bacterial sex

plasmid DNA  carries gene for sex pilus; F+

cell-cell contact required

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Bacterial Growth Conditions
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– Temperature

– Nutrients

– Water (osmotic balance)

– Appropriate atmosphere

– pH

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Bacteriostatic
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stops bacterial growth

-static/-stasis = standing still

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Microbial Death
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D value; decimal reduction time

time require to kill 90% of microbes or spores

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anti microbial targets
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ribosomes

DNA

Enzymes

Cell wall

Membranes

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what affects D-Value
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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

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high temperature physical agents
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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

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Dessication
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freeze dry
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radiation
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physical

DNA damaged

ionzing- gamma rays/xrays

non-ionizing- UV light

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Filtration
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physical

seperates bacteria from liquid

HEPA filtration for air

ultra filtration- filters out lipopolysaccharide endotoxins from dead gram -

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phenolic disinfectant
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chemical

Bactericidal or bacteriostatic depending on 

concentration

higher concentration = bacterialcidal

– Increases membrane permeability

– Denatures protein

 

 

 

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Halogens as disinfectants
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Chemical

Oxidizing agent, inactivates key proteins

 

Iodine – very effective germicide

 Chlorine

 

 

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Heavy Metals as disinfectants
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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
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chemical

Surface active compounds (“surfactants”)

– Damage cell membranes

– Denature protein

 

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Quaternary Ammonium Compounds
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chemical

disinfecting, sanitizing, and antisepsis

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Antibiotics
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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
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Active against Gram positives

– Natural or semi-synthetic

– Allergies and pathogen resistance

ex PENECILLIN 

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Inhibitors of DNA Function
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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
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Enzyme inhibitors

ex. Sulfonamide

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Saprophytic fungi
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decompose organic matter
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Fungi cause Disease in
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• Humans and animals

• Plant pathogens

 • Pests

 

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marine phytoplankton are responsible for
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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
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prokaryotic thus call Cyanobacteria
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Protozoa
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Protozoology: study of protozoa

Eukaryotic, unicellular animals

Motile by cilia, flagella, or amoeboid motility

 

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 Virus Characteristics
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Require host machinery to multiply

1 nucleic acid- DNA or RNA

acellular

capsid- some have lipid envelope

Nucelocapsid

Virion- complete individual virus

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Virus size/shape
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10nm - 300nm

helical- tube shape

polyhedral- multi-sided

 

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lytic bacteriophage infection process
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• 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

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lysogenic phase process
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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
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contain envelopes with Glycoprotein spikes

'naked' virion = non-enveloped

 

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oncogenic viruses
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Incorporation of viral DNA into host DNA may interrupt cell function
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Treatments for Viruses
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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

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