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B12 > Bacterial Transformation (13.2) > Flashcards

Bacterial Transformation (13.2) Flashcards

1
Q

Describe plasmids

A

Small, circular DNA molecules found in bacterial cells

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2
Q

State what plasmids are often used as

A

Vectors

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3
Q

Provide an alternative term to describe vectors

A

Carriers

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4
Q

State the role of plasmids in DNA exchange

A

Employed as vectors to move target DNA from one organism to another.

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5
Q

State what genes can be inserted into

A

Plasmids

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6
Q

State by what process genes from unique organisms can be incorporated into bacterial cells

A

Bacterial transformation

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7
Q

State what the inserted gene can be replicated by

A

Self-replicating properties of plasmid & bacterial cells

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8
Q

State what an inserted gene can express once replicated

A

Proteins for which it codes

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9
Q

Describe restriction enzymes

A

Naturally-occurring bacterial enzyme which can be employed in genetic engineering

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10
Q

Provide an alternative term for restriction enzymes

A

Endonucleases

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11
Q

State what endonucleases have enabled

A
  • cutting of DNA into smaller, more usable fragments

- isolating regions of interest

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12
Q

State what endonucleases compose

A

Bacterial cell’s defence system

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13
Q

State what endonucleases target as part of the bacterial cell’s defence system

A

Foreign DNA that may enter the cell

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14
Q

Describe bacteriophages

A

A virus that infects a bacteria

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15
Q

State what each restriction enzyme generally targets

A

Specific sequence of nucleotides (usually 4-6 base pairs in length)

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16
Q

State what a specific sequence of nucleotides targeted by restriction enzymes is referred to as

A

Recognition site

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17
Q

State what occurs every time a restriction enzyme passes its recognition site

A

Breaks the phosphodiester backbone once on each DNA strand

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18
Q

State whether or not bacterial cells cut up their own DNA

A

No.

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19
Q

State what the name of the enzyme in bacteria which adds a methyl group to a specific nucleotide is

A

Methylase

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20
Q

State where methylase adds a methyl group to a specific nucleotide in bacteria

A

Within the recognition site of the restriction enzymes made by the bacteria

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21
Q

State what adding specific nucleotides within the recognition site of the restriction enzymes made by the bacterium blocks

A

Restriction enzymes from binding to and cutting the baterium’s own DNA

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22
Q

State the 2 types of retstriction enzymes

A
  1. sticky-end restriction

2. blunt-end restriction

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23
Q

State how sticky-end restriction enzymes leave DNA fragments

A

Overhanging ends

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24
Q

State where sticky-end restriction enzymes cut the DNA backbone

A

At different locations on each strand within the recognition site

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25
Q

State what cutting the DNA backbone at different location on each strand within the recognition site results in

A

A staggered cut, leaving ‘sticky ends’

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26
Q

State what the ‘sticky ends’ left by the sticky-end restriction enzyme action are composed of

A

Two fragments have bases exposed

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27
Q

State what the exposed bases left by the ‘sticky end’ restriction enzyme can form

A

Complementary base pairs

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28
Q

State how exposed bases left by the ‘sticky end’ of a restriction enzyme interact

A

Hydrogen bonding occurs between nucleotides of other DNA molecules that have complementary sticky ends

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29
Q

Provide 1 example of a sticky-end restriction enzyme

A

EcoRI

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30
Q

Describe blunt-end restriction enzymes

A

A restriction enzyme that leaves clean-cut ends because it cuts both strands of the DNA molecule at the same location within the recognition site

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31
Q

Describe sticky-end restriction enzymes

A

A type of restriction enzyme that makes a staggered cut in DNA to leave fragments with overhanging ends

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32
Q

State how blunt-end restriction enzymes leave DNA fragments

A

Clean-cut ends

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33
Q

State where blunt-end restriction enzymes cut the DNA backbone

A

At same location on each strand within the recognition site

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34
Q

Provide 1 example of a blunt-end restriction enzyme

A

HaeIII

35
Q

State what restriction enzymes cleave

A

The phosphodiester bonds in double-stranded DNA

36
Q

State what the base-pairing ability of sticky ends allows DNA from different species to do

A

Ligate

37
Q

State what ligation between different species DNA forms

A

Recombinant DNA molecules

38
Q

Describe ligation

A

The process of joining two fragments of DNA using DNA ligase enzymes

39
Q

State what small variations in DNA sequences occur within a population are termed

A

Polymorphisms

40
Q

Describe ligases

A

A group of enzymes that join fragments of DNA or RNA in a process called ligation

41
Q

State what DNA ligase joins

A

Fragments of DNA

42
Q

State what RNA ligase joins

A

Fragments of RNA

43
Q

State the function of DNA ligase

A

Joins fragments of DNA

44
Q

State the function of RNA ligase

A

Joins fragments of RNA

45
Q

State whether or not DNA ligase is also required to join segments of newly replicated DNA and to repair breaks in DNA molecules

A

Yes

46
Q

State what bond ligases forms between two DNA fragments

A

Phosphodiester bond

47
Q

State whether or not ligation of sticky-end fragments is specific

A

Yes.

48
Q

State why the ligation of sticky-end fragments is specific

A

Exposed bases of sticky-end fragments first bind to complementary bases

49
Q

State whether or not ligation of blunt-end fragments is specific

A

No.

50
Q

State why the ligation of blunt-end fragments is random

A

Any two fragments can join if they come in contact and DNA ligase is present to join them

51
Q

State why blunt-end fragments are difficult to use in DNA manipulation processes

A

Ligation is random

52
Q

State whether or not plasmids replicate independently of the chromosome

A

Yes. Plasmids replicate independently of the choromosome.

53
Q

State what is formed when DNA from two different sources is joined

A

Recombinant DNA

54
Q

State why scientists create recombinant DNA

A

To clone particular genes

55
Q

State what scientists commonly use as a vector when creating recombinant DNA

A

Plasmids

56
Q

State what scientists insert into the plasmid to produce a recombinant plasmid

A

Target DNA

57
Q

State where the recombinant plasmid is placed after formation

A

Bacterial cell

58
Q

State what occurs when the recombinant plasmid is introduced back into the bacterial cell

A

Self-replicating system of the plasmid & cell replicates plasmid genes

59
Q

State 3 reasons supporting the use of plasmids as vectors when creating recombinant DNA

A
  1. small size (ease of manipulation)
  2. carry range of restriction enzymes
  3. self-replicate independently once placed in host bacterial cell
60
Q

State whether or not recombinant plasmid self-replicate at a faster rate than the bacterial’s host chromosomal DNA

A

Yes.

61
Q

State 2 factors required of plasmids for the identification of cells that have incorporated the recombinant plasmid

A

Plasmids need:

  1. antibiotic resistant gene
  2. gene that can be easily identified
62
Q

Describe reporter gene

A

A gene that allows detection of gene expression in genetic engineering

63
Q

State the 4 steps involved in the process of creating recombinant plasmids

A
  1. Cut target DNA using sticky-end restriction enzyme and isolate
  2. Cut bacterial plasmid by same restriction enzyme
  3. Place target DNA and plasmids together
  4. DNA ligase added to rejoin backbone
64
Q

State what DNA is employed as target DNA to produce a eukaryotic protein in a bacterial cell

A

Complementary DNA

65
Q

Describe cDNA

A

DNA that has been copied from mature mRNA and contains only exons

66
Q

State how cDNA is synthesised

A

Using reverse transcriptase enzyme

67
Q

State what reverse transcriptase has the ability to make cDNA from

A

mRNA

68
Q

State why the ability of reverse transcriptase to make cDNA from mRNA is useful

A

Mature mRNA has already had the introns spliced out

69
Q

State whether or not prokaryotic cells are unable to splice out introns

A

Yes.

70
Q

Reverse transcriptase allows the synthesis of DNA from what?

A

RNA in a test tube

71
Q

State what occurs when cDNA is inserted into a plasmid and thus, incorporated into a bacterial cell

A

Protein encoded by cDNA will be expressed

72
Q

State for what purpose regulatory genes may be included in plasmids

A

Purpose of controlling the expression of the target gene that is inserted into the plasmid

73
Q

State what the regulatory gene is turned on by

A

An inducer molecule

74
Q

State what occurs once the regulator is transcribed

A

The target gene can be transcribed and translated

75
Q

State what inducers are important in

A

Regulating gene expression

76
Q

Describe inducers

A

A molecule that regulates gene expression

77
Q

State what cells that have foreign DNA introduced to them are called

A

Transformed cells

78
Q

State the 2 forms of artifical bacterial transformation

A
  1. Heat shock

2. Electroporation

79
Q

Describe heat shock

A

Involves placing bacterial cells and a mixture of recombinant and non-recombinant plasmids in an ice-cold solution

80
Q

Describe electroporation

A

Bacterial cells and a mixture of recombinant and non-recombinant plasmids are subjected to an electrical current that alters the plasma membrane

81
Q

State, after heat shock and electroporation, whether or not many bacterial cells will be transformed with recombinant plasmids.

A

No. Very few.

82
Q

State what genes all plasmid vectors contain

A
  • gene for antibiotic resistance

- gene that displays a particular phenotype

83
Q

State how it can be determined whether or not bacterial cells have been transformed

A

Cells are incubated at 37C upon agar plates that contain antibiotic
Those that survive will be those that have taken up the plasmid

84
Q

Describe genetic transformation

A

Process that involves the introduction and expression of foreign genes in a host organism.

B12 (41 decks)

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