Brainscape's Knowledge GenomeTM

Browse over 1 million classes created by top students, professors, publishers, and experts.


See full index

Biomedical Lab > Lab 6 > Flashcards

Lab 6 Flashcards

1
Q

Competent e.coli bacterial cells are prepared using….

A

Calcium chloride

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

The competent E.coli are transformed with the _____ ______ ___ using ______

A

PGLO plasmid DNA using heat shock

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

Non transformed AND transformed E.coli are grown on similar…? What does this detect?

A

Grown on similar agar plates. To detect the phenotypic differences

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

The E.coli colonies on the various agar plates will be evaluated for what 2 things?

A

Antibiotic resistance and fluorescent phenotypes

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

What 2 things are isolated and tested to understand the relationship between genotype, gene regulation, and phenotype?

A

PGLO plasmid DNA and GFP (Green fluorescent protein)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

Each student will use PGLO transformed E.coli culture grown using ____ or ____

A

Antibiotic selection or Antibiotic selection AND AraC gene regulation

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

What are prokaryotes?

A

Organisms made up of cells that lack a cell nucleus or any membrane-encased organelles

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

Give an example of a prokaryote.

A

Bacteria

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

What are PLASMIDS?

A

Small, circular extrachromosomal DNA found in bacteria

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

Plasmids replicate _____ of the chromosome and generally result in ______ changes required for survival in a hostile environment

A

Plasmids regulate INDEPENDENTLY of the chromosomes and generally result in PHENOTYPIC changes required for survival in a hostile environment

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

Give an example of when a plasmid would replicate in attempt to survive in a hostile environment

A

Antibiotic resistance

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

Plasmid DNA transfer occurs_____ in bacteria through a process known as _____

A

Occurs naturally, transformation

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

Transformation is used in……

A

Recombinant DNA technology

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

Can transformation be performed in a lab?

A

YES easily

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

What is the genome?

A

Complete set of genetic information in an organism

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

How is the genome stored?

A

In the form of long DNA molecules called chromosomes

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
17
Q

What are genes?

A

Short segments of DNA containing the protein coding information.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
18
Q

Where are genes located (don’t say nucleus)

A

In the chromosomal DNA

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
19
Q

In higher organisms, where else are genes present besides chromosomal DNA?

A

Mitochondrial DNA

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
20
Q

DNA stands for…..

A

Deoxyribonucleic acid

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
21
Q

DNA molecules are very long chains of repeated subunits known as…

A

Nucleotides or bases (A,G,C,T)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
22
Q

DNA strands are…

A

Antiparallel

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
23
Q

The 2 DNA strands are twisted to form a ——

A

Helix

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
24
Q

DNA is also present in which 2 places?

A

In eukaryotic mitochondria and in bacterial plasmids

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
25
Q

Mitochondrial and plasma DNA are called…?

A

Extrachromosomal DNA

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
26
Q

The antiparallel strand reads __to___

A

3 prime-5 prime

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
27
Q

The parallel strand reads

A

5 prime-3prime

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
28
Q

DNA is said to be____ed

A

Supercoiled

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
29
Q

PGLO contains several ___ and ____ that enable ____

A

Genes and DNA sequences that enable replication of the plasmid DNA

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
30
Q

What is GFP?

A

The jellyfish gene that encodes green fluorescent protein

31
Q

What is responsible for the green fluorescent phenotype?

A

GFP gene

32
Q

What is ori?

A

The ORIGIN of pGLO plasmid DNA replication

33
Q

What is ampicillin?

A

An antibiotic that kills bacteria

34
Q

How is ampicillin destroyed?

A

By the enzyme beta-lactamase (encoded by bla gene)

35
Q

Do TRANSFORMED BACTERIA die in the presence of ampicillin?

A

NO-THEY SURVIVE

36
Q

What is bla?

A

A gene that encodes the enzyme beta-lactamase. Responsible for antibiotic resistant phenotype.

37
Q

Does bla allow bacteria to grow in the presence of ampicillin?

A

Yes

38
Q

What does the enzyme beta lactamase do and what gene encodes it?

A

Breaks down ampicillin. Encoded by bla gene

39
Q

What is the pBAD promoter?

A

Specific DNA sequence upstream from GFP gene.
Binds araC-arabinose and promotes RNA polymerase binding and transcription of GFP

40
Q

What does the araC gene encode?

A

Encodes the regulatory protein that binds to the pBAD promoter

41
Q

When is the production of GFP switched on?

A

ONLY when arabinose binds to the araC protein

42
Q

What binds to the pBAD promoter?

A

The araC protein

43
Q

The araC protein is encoded by…

A

The araC gene

44
Q

What is arabinose?

A

A sugar that binds to the pBAD promoter and displaces the AraC protein from it so RNA polymerase can bind and initiate transcription

45
Q

How is recombinant DNA formed?

A

DNA from 1 organism foreign to the bacteria (or yeast) is inserted into their plasmid

46
Q

What is a DNA VECTOR?

A

The recombinant plasmid formed from inserting a foreign piece of DNA into bacteria

47
Q

The recombinant plasmid is transformed in…?

A

Cells

48
Q

Recombinant plasmid is transformed in cells. The cells are grown in media to produce more ____ or _____

A

Plasmids or the gene product (proteins)

49
Q

Recombinant DNA technology is useful in..?

A

Preparing proteins that are used in research and therapy

50
Q

Recombinant DNA technology involves 4 steps:

A
  1. Synthesis of recombinant plasmid
  2. Transformation of the organism
  3. Selection of the transformed bacteria
  4. Synthesizing the gene product (protein of interest)
51
Q

“Synthesizing a recombinant plasmid”
What MUST the plasmid contain?

A

-Gene of interest (duh)
-DNA for replication (origin of replication)
-antibiotic resistance gene for survival in an antibiotic containing media

52
Q

What is an optional thing for the plasmid to contain?

A

Other genes for regulating the transcription of the proteins

53
Q

“Transformation of the organism”
What does this mean?

A

The plasmid is inserted in a bacterial cell

54
Q

“Selection of the transformed bacteria..”
What does this mean?

A

Transformed cell is grown in an antibiotic containing media. Therefore, only the bacterial cells containing the plasmid will grow. Non transformed cells will die

55
Q

“Synthesizing the gene product(protein)”
How is this done?

A

The cells are grown in media that contain molecules required for producing the protein

56
Q

What does it mean to be a competent cell?

A

The cells are capable of taking in foreign DNA

57
Q

What are cells treated with to make them competent?

A

Calcium chloride (CaCl2)

58
Q

How does calcium chloride enable cells to be competent?

A

The positive charge of calcium shields the negative charge of DNA phosphates. Makes it easy for DNA to enter the cell through the cell membrane

59
Q

How are competent cells stored before they are used?

A

They are frozen

60
Q

While competent cells are kept frozen until use, what prevents the formation of ice crystals and prevents them from dying?

A

Glycerol

61
Q

Transformation is a method to introduce ____ into _____

A

Introduce plasmid DNA into bacteria

62
Q

DNA is very ____ charged

A

Negatively

63
Q

After the bacterial cells uptake the foreign DNA, then what?

A

Bacteria are shocked using hot/cold treatment. Ice slows fluidity of the membrane. Then, the transformation mixture is quickly warmed up. Shocks the cells and increases permeability of the cell membranes

64
Q

How are shocked cells recovered?

A

Kept in nutrient broth for a short period of time. Recovered cell suspension is grown overnight on agar plates

65
Q

Each colony represents…

A

A single transformed cell (clone)

66
Q

Plates without antibiotic are used as…

A

Controls to demonstrate that the cells were alive and survived the transformation

67
Q

Agar plates WITH antibiotic are used to…

A

Select the transformed cells. These cells are the antibiotic resistant phenotype and grow in the presence of the antibiotic

68
Q

What is RNase? To what buffer is it added and why?

A

Enzyme that rapidly degrades RNA. RNASE is added to P1 buffer to exclude the RNA in our DNA extract

69
Q

BUFFER P2
solution that appears clear shows the ____ of cells and _____ of cellular components

A

lysis, solubilization

70
Q

the cell suspension will turn ___after the addition of P2

A

blue

71
Q

BUFFER P1
solution that appears cloudy shows the ____of cells

A

suspension

72
Q

which buffer is the neutralization buffer?

A

N3

73
Q

in N3, solution changes from blue to…

A

colorless

74
Q

in N3, the solution appears as a milky precipitate. why?

A

insoluble DNA and proteins

Biomedical Lab (10 decks)

Brainscape helps you reach your goals faster, through stronger study habits.
© 2024 Bold Learning Solutions. Terms and Conditions