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Microbiology > Quiz 4 > Flashcards

Quiz 4 Flashcards

Quiz 4 transformations and media types

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Biology 101 flashcards
1
Q

MacConkey Agar (MAC)

A
  • Selective and differential
  • Useful in ID of Enterobacteriaceae (Gram-)
  • Clinical laboratories
  • Bile Salts, Crystal Violet: Inhibits Gram+
  • Neutral Red dye: Stains microbes that ferment lactose. Ph is lowered so they get stained a pink color.
  • Escherichia coli will grow red/pink colonies because it will use the lactose in the media.
  • Non-fermenters will grom white/colorless because they used peptoe instead of lactose. This produces ammonia.
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2
Q

Eosin Methylene Blue Agar (EMB)

A
  • Selective and Differential
  • Inhibits Gram+ and Selects Gram-
  • differentiates coliform and non-coliform GNRs
  • Used to test quality of water. Not used in a lab setting
  • Eosin Y Methylene blue: Ph indicator forms dark purple precipitate at low Ph due to fermentation
  • Lactose: Coliform will ferment the lactose
  • Escherichia coli has a strong fermentation and precipitation of acid and indicator complex. This will case it to produce a green metallic sheen.
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3
Q

Coliform

A
  • GNR and ferment lactose
  • Escherichia coli
  • Enterobacter
  • Serratia
  • Citrobacter
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4
Q

CHROMOGENIC MEDIA

A
  • ID of certain bacteria
  • Chromogens: convert to different colors when degraded by different microbe’s enzymes
  • Rapid detection of MRSA
  • New ones created for drug resistant pathogens like Escherichia coli, Candida
  • HardyCHROM UTI: used for urinary tract pathogens
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5
Q

FERMENTATION AND UTILIZATION MEDIA

A

Help identify bacteria based on:

  • Sugar they utilize for fermantiation
  • the acids they produce
  • Ability to utilize O2
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6
Q

PHENOL RED BROTHS

(Durham Tube Sugar Fermantations)

DEXTROSE, LACTOSE, SUCROSE (GNR)

A
  • Distiguishes carbohydrate fermenters from non fermenters.
  • Distiguishes which carbohydrate was used by the products formed
  • Media: 0.5-1% Carb broth. Dextrose, Lactose or Sucrose
  • Pepton with Phenol red: The inverted Durham tube for gas detection
  • Carb fermentation results in Acid. Those may also produce Gas.
  • Gas production= gas bubbles in Durham tube
  • Acid= Color changed form red to yellow
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7
Q

METHYL RED TEST (MR)

Mixed Acid Fermentation

A

Determines mixed acid fermentation

Part of IMVic tests

MRVP broth: Media of pepto glucose broth used for this test and the VP test

Methynol Red: Added after incubation for pH indication

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8
Q

VOGES PROSKAUER TEST (VP)

Butanediol Production

A
  • Detects production of acetoin or butaneidol from fermantation of glucose
  • part of IMVic test
  • MRVP media: glucose pepto
  • Barrit’s reagents A (naphthol) added after incubation
  • Barrit’s reagents B (potassium hydroxide, KOH) added after incubation
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9
Q

CATALASE TEST

A
  • Detects the production of the anzyme Catalase
  • TSA media
  • Reagent: 3% Hydrogen Peroxide (H2O2)
  • Hydrogen Peroxide is a by-product of oxidative repiration. Catalase converts Hydrogen Peroxide to Oxygen and Water. Anaerobes lack this enzyme
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10
Q

OXIDASE TEST

A
  • Detects the production of enzyme cytochrome or oxidase
  • We used oxidase dry slides
  • Organisms that use oxygen will have the cytochrome oxidase transfers electrons to the oxygen.
  • Thre reagens in dry slide will change from yellow to purple when oxidized
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11
Q

CITRATE

A

Determine ability to use citrate as sole source of carbon.

Part of IMVic tests

Media: Simmons Citrate Agar. Contains sodium citrate as sole carbon source, mineral salts and pH Bromothymol blue.

Bromothymol blue: pH indicator

Utilization of citrate leaves a sodium residue. This increases pH

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12
Q

GNR

A
  • Escherichia coli
  • Serratia marcescens
  • Pseudomonas fluorescens
  • Enterobacter aerogenes
  • Alcaligenes viscolactis
  • Citrobacter freundii
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13
Q

LYSES

A

Rupture of cell wall of membrane

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14
Q

LYTIC

A

It causes lysis

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15
Q

LYTIC CYCLE

A

Two basic phage infection cycles:

  • Lytic cycle
  • Lysogenic cycle
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16
Q

SOFT AGAR OVERLAY

A
  • 1% Agar
  • Soft agar in a tub, kept in hot water bath
17
Q

CONFLUENT LAWA OF GROWTH

A

When the growth of bacteria covers entire surface of media

18
Q

PLAQUES

A
  • Space where the bacteria have been lysed by the bacteriaphage, their cells are broken down and no longer visible.
  • Clear spots
19
Q

TRAMSFORMATION

A

When the bacteria takes plasmid DNA from the surrounding environent.

20
Q

CONJUGATION

A

Bacteria transfer genes from one bacterium to another

21
Q

GENE REGULATION

A

Expressing the gene that was obtained through transformation

22
Q

GENETIC ENGINEERING

A

Forcing a bacterium to take in a plasmid DNA from the surrounding and express that gene. It would not normally have that gene.

23
Q

pAMP

A

Plasmind that contains a gene that provided Escherichia coli with Ampicillin resistance

24
Q

RECOMBINANT PLASMIDS

A
  • Plasmids constructed form genes of different organisms
  • In a lab, plasmids can be combined with other DNA plasmids.
  • For example, combining antibiotic resitant gene with another gene
25
Q

SELECTALE MARKERS

A

Used to help us differentiate the bacteria that were transformed. If we add a recombinan plasmid of a gene that allos a bacteria to be resistant to antibiotic and another gene of interest. The bacteria that groes even in the prescence of antibiotic would be the “selectable marker” because those are the bacteria that have taken both genes.

26
Q
A
27
Q

CAPSULE

A
  • One type of glycocalyx

Structure present in some bacteria:

  • Protects cell from dehydration and nutrient loss
  • Inhibits killing by WBC through phagocytosis
  • Aids bacteria in attachment (biofilms)
28
Q

PLASMIDS

A
  • Nonessential pieces of DNA seperated from chromosomes
  • ofter confer protective traits suck as resistance to antibiotics
  • Easily manipulated in a lab setting which makes them important in modern engineering techniques.
29
Q

GFP

A

Green fluorescent protein pGLO

30
Q

RECOMBINANT pGLO PLASMID

A

Three different gens joined together

Bla: betalactamese - makes bacteria resistant to beta-lactam antibiotics (Penicillin)

graC: Regulates expression of GFP. (expressed only in arabinose)

31
Q

COMPETENT CELL

A
  • Cells capeble of undergoing transformation by using special DNA binding receptors on their cell wall
  • Cells whose cell wall can be manipulated into allowing DNA to pass through
  • In the lab, we made Escherichia coli cpmpetent by using calcium chloride and heat shock
  • During rapid growth it is easier to make cells competent than in other stages
32
Q

SMOOTH COLONIES (S)

A
  • Strain is virulent
  • Prevents detection by immune system
33
Q

ROUGH COLONIES (R)

A
  • Strain is benign
  • lacks protective capsule
  • Destroyed by immune system
34
Q

ARABINOSE

A

A sugar

In our experiment, it will regulate the expression of GFP

It will only glow in the prescence of this sugar

35
Q

LOG GROWTH

A

Rapid cell division

36
Q

LAG PHASE

A

Very slow growth because cell number is slow.

37
Q
A

Microbiology (12 decks)

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