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Micro5 > Lab Exam 3 > Flashcards

Lab Exam 3 Flashcards

1
Q

What is transformation

A

Type of horizontal gene transfer - recipient cell take up dna from donor

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2
Q

Why is it useful for bacteria to preform transformation

A

Allows bacteria to acquire new traits

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3
Q

How is a bacterial cell made competent in the transformation lab?

A

It is chemically induced with an ice shock and calcium chloride and then a heat shock to let the dna pass through the membrane

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4
Q

What is a vector

A

A plasmid that Carries foreign dna into bacterial cells

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5
Q

Only a small percentage of bacterial cells will take up plasmid dna

TRUE or FALSE

A

True

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6
Q

Recombinant plasmid

A

Plasmid that successfully gets inserted into MCS

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7
Q

What color colonies will a recombinant plasmid be

A

White

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8
Q

MCS (multiple coding site)

A

Where the dna of interest is inserted by researcher

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9
Q

Which plasmid was used in the transformation lab

A

Puc18

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10
Q

What are the 3 important sites we looked at in the transformation lab

A

Lac z gene
Bla gene
Multiple cloning site

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11
Q

The bla gene

A

Allows us to determine which cell has a plasmid

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12
Q

What does the bla gene produce

A

Enzyme penicillinase - breaks down beta lactam ring

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13
Q

What happens once ampicillin has broken the beta lactam ring

A

It can not inhibit the growth of bacteria therfore it will continue to grow

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14
Q

Lac z (b- galacoside) gene

A

Allows us to determine which cell has a recombinant plasmid

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15
Q

What is the purpose of beta galactosidase

A

Metabolizes xgal on the plate

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16
Q

What occurs when the lac z gene (beta galactosidase) metabolizes xgal

A

A blue color is produced

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17
Q

Where is the multiple cloning site (mcs) on the puc18 plasmid

A

In the lac z gene

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18
Q

If new foreign dna is able to be inserted into the multiple cloning site of a plasmid, the plasmid is

A

Recombinant

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19
Q

If the new foreign dna is not able to insert into the multiple cloning site of the plasmid, then the plasmid is

A

Non recombinant

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20
Q

Non recombinant plasmid

A

No dna in the multiple cloning site

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21
Q

What color would a non recombinant plasmid be

A

Blue!! Due to the xgal not being interrupted

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22
Q

Why did we use water as the control in the transformation lab

A

To see if e. Coli will grow

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23
Q

What was in the lb ++ plates used in the transformation lab

A

Xgal
Ampicillin

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24
Q

What is a mutagen?

A

A chemical substance or physical agent that can induce changes in the base sequence of dna

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25
Q

What does salmonella need in order to grow?

A

Histidine

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26
Q

Histidine

A

Used by bacteria to grow and survive

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27
Q

What type of media plate does not contain histidine

A

Minimal media

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28
Q

Purpose of the Ames test

A

To show rather or not a certain chemical substance is a mutagen

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29
Q

Back mutagen

A

Mutation that reverts the bacteria genome to a while type (Normal)

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30
Q

Wild type

A

What is found in nature ( normal type)

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31
Q

What type of mutation is present in salmonella F in the Ames test?

A

Frameshift mutation

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32
Q

What type of mutation was present in the salmonella M strain?

A

Base pair mutation

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33
Q

What was the negative control plate used in the Ames test?

A

Minimal media plate with disc soaked in water

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34
Q

What is the outcome for the negative control plate possibly having colonies? _ the water

A

Spontaneous mutation occurring during dna replication allowing salmonella to grow w/o histidine

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35
Q

What was the positive control plate in the Ames test?

A

Minimal media plate w/ disc soaked in NaAz

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36
Q

What is sodium azide (NaAz) and what does it cause?

A

It is a mutagen
- causes: base pair mutation

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37
Q

4-Nitro is? And causes what?

A

Mutagen that causes a frameshift insertion mutation

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38
Q

4- nitro is capable of fixing what salmonella strain

A

F

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39
Q

NaAz is capable of fixing what salmonella strain

A

M

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40
Q

What would a positive control plate look like in the Ames test?

A

Growth of salmonella due to presence of known mutagen

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41
Q

What does no growth indicate on a minimal media plate during the Ames test

A

The chemical is not a mutagen

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42
Q

What type of plate was used in the toxicity test?

A

Complete media

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43
Q

What does complete media contain that minimal media doses not?

A

Histidine

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44
Q

If a zone of inhibition is present during a toxicity test that indicates

A

The chemical is toxic to the bacteria therfore they die which causes the clearing

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45
Q

What are the 6 metabolic tests used to identify bacterial species

A

Catalase
Coagulase
Mannitol salt agar
Triple sugar iron test
Coli form test
Entertube test

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46
Q

What type of bacteria does the catalase test identify?

A

Bacteria that can preform aerobic respiration

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47
Q

What type of bacteria does the coagulase and mannitol salt agar test identify?

A

Staphylococcus species

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48
Q

What type of bacteria does the coliform test identify?

A

Bacteria, usually e. Coli present in water as fecal contamination

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49
Q

What type of bacteria does the triple sugar iron test identify ?

A

Enteric bacteria

50
Q

What is catalase

A

An enzyme that breaks down hydrogen peroxide to water and oxygen

51
Q

Why would a bacteria need to produce catalase?

A

To breakdown hydrogen peroxide since it is toxic as byproduct in aerobic respiration

52
Q

What type of bacteria needs catalase

A

Ones that can preform aerobic respiration

53
Q

What type of bacteria preform anaerobic respiration

A

Obligate aerobes
Facultative anaerobes

54
Q

If a bacteria did not produce bubbles in the catalase lab what does that mean?

A

Catalase reaction did not occur

55
Q

What locations on the body might you find bacteria with the catalase enzyme?

A

Mouth or skin

56
Q

Where on the body might you find catalase enzyme not present?

A

Intestines

57
Q

Difference between fermentation and anaerobic respiration

A

Electron transport chain not present in fermentation

58
Q

What is coagulase?

A

Enzyme that catalyzes conversion of fibrinogen into fibrin

59
Q

Why does a bacteria need to produce coagulase

A

Helps to evade hosts immune system

60
Q

How does clotting the blood help. A bacteria envied the host’s immune system?

A

Fibrin creates a protective barrier by surrounding saureus cells protecting it from immune cells which will kill them

61
Q

Why is coagulase considered a virulence factor?

A

Causes bacteria to survive and continue to replicate in the host cell

62
Q

What type of blood plasma did we us in the coagulase test in lab?

A

Rabbit plasma

63
Q

What would a positive coagulase test look like? negative?

A

Positive : clot formed - thick and cloudy

Negative : no clot - liquid

64
Q

What type of media is mannitol salt agar?

A

Selective - halo tolerant bacteria

Differential - staphylococcus aureus

65
Q

What ingredient in MSA plates make it selective?

A

7.5% sodium chloride
Ph indicator

66
Q

How is MSA plates differential?

A

They distinguish between bacteria that can ferment and those that can not

67
Q

What ingredients in a MSA plate make it differential?

A

Mannitol
Ph red indicator

68
Q

Mannitol is

A

A carbohydrate broken down via fermentation

69
Q

What is the important by product of fermentation of mannitol?

A

Lactic acid

70
Q

What does the phenol red indicator measure in the MSA test?

A

Presence of lactic acid indicating bacteria can do fermentation

71
Q

If phenol red indicator turns yellow, what does that mean

A

Ph level = low
- lots of lactic acid therefore can ferment mannitol

72
Q

If the phenol red indicator turns pink on the MSA plate, what does that mean?

A

Ph is high
- no lactic acid therefore an not ferment mannitol

73
Q

MSA -/- plate

A

Can not grow in high salt environment and can not breakdown mannitol to lactic acid

74
Q

MSA +/+ plate

A

Can grow in high salt environment and can breakdown mannitol
- will be yellow

75
Q

MSA +/- plate

A

Can grow in high salt environments and can breakdown mannitol but can not break down to lactic acid (pink)

76
Q

Enteric bacteria are gram+ or gram-?

A

Gram -

77
Q

Enteric bacteria are coccus or bacillus?

A

Coccus

78
Q

Are enteric bacteria facultative aerobes or facultative anaerobes?

A

Facultative anaerobes

79
Q

What are the two abilities of enteric bacteria?

A

Able to ferment sugars and breakdown amino acids

80
Q

What is the triple sugar iron test used for?

A

To distinguish between enteric bacteria

81
Q

What makes up a TSI agar plate

A

Peptone base
Glucose
Sucrose
Lactose

82
Q

Why are glucose, sucrose and lactose used in the triple sugar iron test?

A

To see if the bacteria can ferment them

83
Q

What sugar does bacteria prefer to use the most?

A

Glucose!!!

84
Q

What ingredient allows us to see if a bacteria can ferment any of the 3 types of sugar?

A

The red phenol indicator

85
Q

What does the red phenol indicator measure in the TSI test?

A

Acid by product being used for fermentation

86
Q

How can you tell gas was produced via fermentation?

A

Bubbles occurred

87
Q

What occurs on the slant of the tube?

A

Aerobic respiration

88
Q

What occurs in the butt of the tube in a TSI test?

A

Fermentation

89
Q

No fermentation of sugars has what color slant/butt

A

Slant = pink
Butt = red

90
Q

If a bacteria can only ferment glucose what color will the slant/butt be

A

Slant = pink

Butt = yellow

91
Q

If a bacteria can ferment glucose, lactose and sucrose, what color will the slant/butt be?

A

Slant = yellow

Butt = yellow

92
Q

If a bacteria is able to metabolize cysteine, what color will the slant/butt be?

A

Slant = yellow

Butt = black precipitate

93
Q

What ingredient is added to the TSI test to see if the bacteria can break down the amino acid cysteine?

A

Feso4

94
Q

Obligate aerobe tube in TSI test will have

A

Red slant w/ red butt

  • NO FERMENTATION
95
Q

What does cysteine (amino acid) get broken down to?

A

Hydrogen sulfide

96
Q

The ph indicator in the TSI test detected the breakdown of what amino acid

A

Cysteine

97
Q

What does the coliform Analysis test for?

A

Fecal contamination in water

98
Q

Coliform means

A

Having E. Coli like characteristics

99
Q

E. Coli characteristics

A

Gram-
Bacillus
Can ferment lactose

100
Q

What are the three steps (tests) in Coliform analysis?

A
  1. Presumptive test
  2. Confirmed test
  3. Completed test
101
Q

What type of tube is used in the presumptive test?

A

Lactose broth with small Durham tube inside

102
Q

Why is lactose broth used in the coliform analysis lab?

A

To make sure it selects e.coli - it is needed to ferment

103
Q

The purpose of the first test - presumptive test?

A

To select bacteria that can ferment lactose

104
Q

What type of tube was used in the 2nd test - confirmed test?

A

Brilliant green lactose bile broth w/ Durham

105
Q

What makes the brilliant green lactose broth selective?

A

Brilliant green dye and bile salts

106
Q

How does the brilliant green dye and bile salts make it selective?

A

Inhibits growth of gram+ making it promote the growth of gram-

107
Q

What was the purpose of the 2nd test - confirmed test?

A

To select for bacteria that can ferment lactose and are gram negative

108
Q

What type of plate was used in the completed test in coliform analysis?

A

EMB - eosin methylene blue agar plates

109
Q

what type of media was the methylene blue agar (EMB) ?

A

Selective & differential

110
Q

What made EMB plates selective?

A

The methylene blue inhibited the growth of gram+

111
Q

What made the EMB plates differential?

A

Eosin dye - making the copies look metallic green due to high acid production

112
Q

Whatwas the purpose of the last - completed test in the coliform analysis?

A

To select gram negative bacteria that can ferment lactose by producing lots of acid

113
Q

Does a presumptive test indicate evidence of fecal contamination in a water sample?

A

No- only the presumptive test tells if a bacteria can ferment lactose not if it is gram -

114
Q

Does a confirmed test indicate definitive evidence of fecal contamination in a water sample?

A

No- further testing is needed

  • it confirms the test is positive for fermenting lactose and being gram- but not how well it can ferment
115
Q

Carcinogen

A

Chemical substance or physical agent that is capable of causing cancer

116
Q

What type of bacteria (group) was being tested in the rapid one lab?

A

Enteric bacteria

117
Q

What types of breakdown can enteric bacteria do?

A

Ferment sugar, breakdown amino acids

118
Q

Name a sugar that was used in rapid one

A

Sorbitol , adonitol

119
Q

Name an example of an amino acid in rapid one lab

A

Lysine, arginine

  • ends in ine
120
Q

Name an example of another molecule used in rapid one

A

Urea, fatty acids

121
Q

How could you tell a bacteria was able to ferment sugar, breakdown an amino acid or a molecule in rapid one?

A

Color change occurred - indicating ph change

122
Q

When sugars are broken down in rapid one, does the product become more acidic or basic?

A

Acidic - ph lowers

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