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CBG Bacterial genetics > transformation > Flashcards

transformation Flashcards

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Biology 101 flashcards
1
Q

what is this the definition for? a virus infects a bacterium and some bacterial DNA is entrapped in the viral capside; this virus infects another bacterium and transfers the DNA of the previous bacterium

A

transduction

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2
Q

what is transformation?

A

the bacteria gains a trait by taking in DNA from its environment

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3
Q

what is conjugation?

A

Transfer of DNA from one bacterium (F+) to other (F-) through sex pilus and with the help of F factor plasmid is

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4
Q

are bacteria diploid?

A

no

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5
Q

what is the impact of bacteria being haploid? are they easy to grow?

A

yes easy to grow and test traits, also show no dominance or recessivity

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6
Q

how is there bacterial variation?

A

from errors e.g DNAP

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7
Q

what is an auxotroph?

A

a mutant organism, especially a microorganism, that has a nutritional requirement not shared by the parent organism. (it cannot synthesize compound needed for growth)

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8
Q

what is a prototroph?

A

an organism able to grow and survive on minimum conditions (it provides its own compounds needed for growth)

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9
Q

when did Griffith perform his experiments?

A

1928

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10
Q

what were griffiths observations?

A

virulen strain=smooth non virulent=rough then in mice experiments mix of heat killed smooth and non virulent still caused mice to die –> discovered transformation

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11
Q

what did Frederick Griffith discover in 1928

A

transformation

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12
Q

what did Avery,Macleod and McCarty demonstrate?

A

they showed the ‘transforming principle’ was DNA

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13
Q

how did Avery,Macleod and McCArty demonstrate DNA was the transforming principle?

A

added RNAse,protease and DNAase to the same mix(heat killed virulent and non virulent strain) and only the one without DNA stayed non virulent

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14
Q

in transformation what type of DNA attached onto the cell and what type is taken up?

A

dsDNA binds to cell,endonucleases mean only ssDNA is actually moved into the cell and this ssDNA is inserted into the bacterial chromosome by recombination

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15
Q

what 3 problems can occur in transformation?

A

1)dsDNA may not attach to cell 2)ssDNA may not be taken up into cell 3)ssDNA may not be incorporated into the chromosome by recombination

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16
Q

which process allows ssDNA to get into the bacterial chromosome in transformation?

A

recombination

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17
Q

in transformation is the cell the info is being picked up from dead?

A

yes

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18
Q

why undergo transformation? (3)

A

nutrition,repair and variety (adaptation)

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19
Q

if two donor genes are located closely on the chromosome what is there a greater chance of?

A

they will be carried on the same piece of transforming dna and therefore greater chance of double transformation

20
Q

what can transformation be used for?

A

to assess linkage (test for a departure from the product rule)

21
Q

in transformation does the donor DNA replace a large percent of the recipients chromosome?

A

no, only a small percent (cotransformation is because thE genes are in close proximity to one another)

22
Q

What are PLQ?

A

pores that allow DNA to reach import system

23
Q

what is ComEA protein responsible for?

A

reception of dsDNA on cell surface which is then nucked by endonucleases to become ssDNA

24
Q

what do endonucleases in a competent bacterial cell do?

A

nick the dsDNA to make it ssDNA

25
Q

what is the ssDANA transported through?

A

ComE

26
Q

what can lead to the COM system developing?

A

stress and high density

27
Q

why do bacteria undergo transformation?

A

nutrition,repair,variety

28
Q

what is the pillus function?

A

no known function,perhaps involved in conjugation or pulling up DNA as it contracts and retracts

29
Q

what are two methods of artificial transformation?

A

cacl2 transformation and electroporation

30
Q

what is competence?

A

competence refers to the state of being able to take up exogenous DNA from the environment

31
Q

what system does natural transformation involve?

A

COM system

32
Q

what does electroporation do?

A

artificial transformation-electric field 10-20kV/cm changes permeability of bacterial membranes

33
Q

what voltage is used in artificial electroporation?

A

10-20kV/cm

34
Q

is artificial more or less efficient than natural?

A

less

35
Q

explain Cacl2 transformation briefly?

A

add CaCl2 which promotes binding of plasmid to LPS and calcium ions attract negative DNA. The plasmid DNA can then enter cells after Heat shock (changes membrane permeability) e.g from 4 degrees to 42 for e.coli

36
Q

what is required in CaCL2 transformation to allow plasmid DNA to enter? at what temps?

A

heat shock from 4 to 42 for e.coli

37
Q

what does the Calcium ion in calcium chloride do in artificial transformation?

A

promotes plasmid binding to Lippopolysacharide and the negative DNA is attracted to it

38
Q

what is a polylinker?

A

area for inserting foreign DNA (unique restriction sites are found in this region)

39
Q

where are unique restriction sites found?

A

polylinker region

40
Q

T or F: the rate of cotransformation of 2 genes is inversely proportional to the distance between them?

A

True. The quicker the transformation the closer they are.

41
Q

what is b galactosidase encoded by?

A

lacZ

42
Q

in blue white screening do both e.coli and the plasmid have a mutant lacZ?

A

no, e.coli has mutant lacZ(deletion mutant) and the plasmid has a functional lacZ

43
Q

what does the lacZ in the plasmid have? (blue/white screening)

A

A MCS (multi cloning site)

44
Q

what can MCS in plasmid be cut by? (blue/white screening)

A

restriction enzymes

45
Q

will blue colonies have an inserted vector?

A

no, blue colonies have vector with uninterrupted lacZalpha therefore have no insert

46
Q

in blue/white screening what are the cells grown in the presence of?

A

Xgal

CBG Bacterial genetics (4 decks)

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