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Cell Bio > Biotechnology > Flashcards

Biotechnology Flashcards

1
Q

Many bacteria and certain fungi contain self-replicating extrachomosomal DNA molecules called

A

Plasmids

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2
Q

Plasmid DNA is

A

Closed circular and double-stranded

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3
Q

Self-replicating and do not integrate into the host genome

A

Plasmids

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4
Q

The smallest plasmids

A

contain only a few thousand base pairs

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5
Q

The largest plasmids

A

contain over 100,000 base pairs

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6
Q

Confer a specialized and sometimes protective characteristic to the organism

A

Genetic information for the translation of proteins found in bacterial plasmids

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7
Q

Certain plasmids carry the gene for the enzyme

A

Beta-lactamase

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8
Q

Catalyzes the decomposition of certain antibiotics such as penicillin and amoxicillin

A

Beta-lactamase

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9
Q

Able to pass from an antibiotic-resistant cell to an antibiotic sensitive cell of the same or another bacterial species, thus rendering the latter resistant

A

Plasmids

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10
Q

Posess an ampicillin resistance gene and a Beta-lactamase gene

A

A plasmid from the strain of Escerichia coli (JM101)

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11
Q

One of the fastest methods to isolate plasmid DNA

A

Single-use columns

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12
Q

Based on the principle of anion exchange chromatography

A

Single-use columns

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13
Q

DNA binds to a modified

A

Silica matrix and is eluted using sterile distilled water

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14
Q

Isolation of Plasmid DNA Materials

A
  1. Pre-Lysis Buffer
  2. Alkaline Lysis Solution
  3. Neutralizing Solution
  4. Glassmilk Spinbuffer
  5. Wash Solution
  6. Sterile Distilled Water
  7. RPM Spin Filters
  8. Catch Tube
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15
Q

To resuspend the cell pellet

A

Add 50 micro liters Pre-Lysis Solution and mix by vortexing or pipetting up and down until completely suspended

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16
Q

A white precipitate will form consisting of

A

Cell membranes, proteins, and chromosomal DNA

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17
Q

To elute plasmid DNA from the Spin Filter,

A

Add 50 micro liters of sterile water and spin for 2 minutes to collect DNA in the bottom of the vial

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18
Q

Many types of bacterial can transfer ________ between individual cells.

A

DNA

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19
Q

Under certain conditions, large portions of chromosomes are transferred to the recipient cell

A

transferred to the recipient cell

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20
Q

Under some condition smaller autonomously replicating circular DNA molecules called _______ are transferred

A

Plasmids

21
Q

Plasmid DNAs do not usually

A

encode information essential for the functioning of the cell

22
Q

Bacteria that lack plasmids

A

function normally

23
Q

Bacteria that possess plasmids

A

may be capable of performing additional functions, as for example showing resistance to specific antibodies

24
Q

Some bacteria display _____________, while others do not

A

Competence

25
Q

Ability to take up DNA at a particular stage in the life cycle

A

Competence

26
Q

It is possible to make E. coli cells competent by treatment with

A

certain reagents

27
Q

When the introduction of foreign DNA into a cell alters the phenotype of the cell, the cell is said to have been

A

Transformed

28
Q

The process of introducing DNA into a cell is known as

A

Transformation

29
Q

Most procedures for the transformation of E. coli are based on

A

The observation that when treated with cold CaCl2 and briefly heat-treated, bacteria could be transformed

30
Q

How CaCL2 confers competence to E. Coli

A

Is unknown, as is the mechanism by which DNA enters the cell

31
Q

A number of factors influence the efficiency of transformation including:

A
  1. The strain of bacteria
  2. The growth medium
  3. The stage of the cell cycle
32
Q

In order to detect cells that have taken up plasmid DNA, it is necessary for the plasmid to contain a

A

Marker

33
Q

A gene that when expressed as RNA or protein, reveals the presence of that gene

A

Marker

34
Q

Frequently markers include ________ that confer resistance to antibiotics to which the cell is ordinarily sensitive

A

genes

35
Q

Purpose of the Transformation of Plasmid lab

A

To demonstrate phenotype changes in bacteria that have been transformed with an antibiotic-resistance gene and a metabolic marker

36
Q

E. coli cells are transformed with

A

The plasmid pBLU

37
Q

Carries genes for two identifiable phenotypes: Amp and lacZ

A

Plasmid pBLU

38
Q

Two identifiable phenotypes

A

Amp and lacZ

39
Q

Responsible for bacterial resistance to Beta-lactam antibiotics

A

Amp gene

40
Q

Codes for Beta-lactamase

A

Amp gene

41
Q

Expresses resistance to the antibiotic ampicillin

A

Beta-lacatamase

42
Q

Widely used reporter gene which cods for Beta-galactosidase

A

lacZ

43
Q

5-bromo-4-chloro-3-indolyl-Beta-galactopyranoside

A

X-gal (lactose analog broken down by the enzyme)

44
Q

Produce a visible blue product

A

The enzyme breaks down the lactose analog X-gal

45
Q

The recipient E. coli strain lacks

A

amp and lacZ (thus transformed cells are converted to an amp and lacZ phenotypes

46
Q

Selective agent

A

Ampicillin

47
Q

Only _________ cells are able to grow on medium containing amipicllin

A

transformed

48
Q

Not a selective marker, but only cells transformed with the plasmid metabolize _________ to produce blue colonies

A

X-gal

49
Q

Luria-Bertania Media

A 
Deionized Water
Bactotryptone
Bacto yeast
NaCL
5M NaOH 
1M HCL

Cell Bio (30 decks)

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