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Cell Bio Lab Test#2 > Lab4: Transformation > Flashcards

Lab4: Transformation Flashcards

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Biology 101 flashcards
1
Q

What is the process of transformation?

A

A bacterium is taking up the foreign DNA and incorporating it into its genome

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2
Q

What is a positive factor of transformation?

A

Increase the survival of the bacteria by acquiring new genetic material

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3
Q

What is a competent bacteria?

A

Bacteria with weakened cell walls capable to take up foreign DNA

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4
Q

What are 4 manners that a bacteria can become competent?

A

-Electro corporation (administering an electric shock to weaken the cell wall)
-Stressed by a low nutrients level
-Temperature shocking
-Presence of calcium chloride

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5
Q

Of what is compose a bacterial chromosome?

A

Single circular chromosome

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6
Q

What is a plasmid?

A

Small, circular pieces of DNA that exist outside the bacterial chromosome

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7
Q

What are to important functions of plasmid?

A

-Very stable
-Replicate independently

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8
Q

What is a vector in biology?

A

A vehicle used to carry foreign DNA into a cell

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9
Q

What is a pG plasmid?

A

Artificial plasmid to insert GFP gene into E.coli

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10
Q

What are the two genes of interest in the experiment?

A

-GFP gene
-AmpR gene

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11
Q

What is the selective marker in the experiment?

A

The AmpR gene

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12
Q

What is the function of the AmpR gene?

A

Confers ampicillin resistance to the bacterium, allowing it to grow on ampicillin-containing medium

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13
Q

What are the origin of the GFP gene? And what is its function?

A

Comes from a jellyfish and make bacteria glow green under UV light

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14
Q

What is the role of AmpR gene as a selective marker?

A

Allows only transformed bacteria (with the plasmid) to survive in the presence of ampicillin

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15
Q

What does PBP stands for?

A

Penicillin-binding protein

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16
Q

What is the role of PBP?

A

Helps the bacteria build their cell walls

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17
Q

What is the key role of Ampicillin?

A

Inhibits PBP, preventing cell wall synthesis, thus bacterial growth

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18
Q

What produces Beta-lactamase?

A

Becterai with AmpR gene

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19
Q

What is the function of Beta-lactamase?

A

Breaks down ampicillin, allowing the bacteria to survive and grow in its presence

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20
Q

Draw a scheme of GFP gene expression

A

GFP gene —- controlled by T7 promoter —- recognized by T7 RNA polymerase

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21
Q

What is the function of T7 RNA polymerase?

A

Binds strongly to the T7 promoter than transcribing viral genes

22
Q

In engineered bacterium the T7 RNA polymerase is linked to what?

A

The lac Operon

23
Q

What normally controls the lac operon?

A

Lactose

24
Q

What is IPTG?

A

-An synthetic substitue for allolactose
-An inducer

25
Q

What is the function of IPTG?

A

-Can bind to the lac repressor
-triggers gene expression

26
Q

Why can IPTG trigger gene expression?

A

Because IPTG is not metabolized by the bacteria

27
Q

What is a recombinant plasmid?

A

Plasmid that successfully incorporated the foreign gene (*In this experiment — GFP gene)

28
Q

What is a non-recombinant plasmid?

A

A plasmid that has released without incorporating the foreign gene

29
Q

True or false: During transformation, there is a mix of recombinant and non-recombinant plasmids is produced?

A

True

30
Q

What is the function of the restriction site?

A

Cut the plasmid and gene of interest

31
Q

Where do the plasmids can make many copies of themselves?

A

At the origin of replication

32
Q

What is the reporter gene of the experiment?

A

GFP gene

33
Q

What does the GFP gene can indicate?

A

Whether foreign gene is being expressed in the bacterium— bacterium will glow green when GFP gene is expressed

34
Q

What is a avantage of using GFP gene?

A

Easy identification

35
Q

Where is located the T7 RNA pol gene?

A

On bacterial chromosome

36
Q

What has been attached to the lac promoter?

A

T7 RNA pol gene

37
Q

What events does the presence of lactose triggers? (3 steps)

A
  • Bacterium make T7 RNA pol — bind to the T7 RNA pol promoter on the plasmid — GFP is expressed in large quantities
38
Q

What is the normal inducer for the lac operon?

A

Allolactose

39
Q

Name the 3 steps of lac operon

A

-inducer binds to the repressor
-falls of the operator
-RNA pol transcribe the gene

40
Q

What is the challenge with eukaryotic gene?

A

Bacteria cannot splice eukaryotic gene to remove introns

41
Q

What is the solution to the challenge of eukaryotic gene?

A

Convert mature mRNA to cDNA — using reverse transcriptase

42
Q

What is the specification of complementary DNA?

A

Lacks introns and can be inserted into bacterial plasmid for expression

43
Q

What is the specification of retroviruses?

A

They have a -RNA (instead of a +RNA)

44
Q

How retroviruses infect a cell?

A

Viral RNA is reverse-transcribed into cDNA then integrates the host genome

45
Q

What is a colony?

A

It’s derived from a single bacterium

46
Q

Is the genetical material of 1 colony is identical?

A

Yes

47
Q

In presents of a recombinant gene, growing a colony represents what?

A

“Cloning” a gene

48
Q

What is the objective of the lab?

A

To insert jellyfish into bacterium to make it glow green when exposed to UV light, in the presence of lactose

49
Q

What is the dependent variable of the GFP lab?

A

Numbers of colonies

50
Q

What is the independent variable of the GFP lab?

A

The presence of plasmid

51
Q

What is the control variable of the GFP lab? And why?

A

IPTG because its concentration remains constant

52
Q

What is the special treatment given to E.coli in this experiment?

A

Add of calcium chloride (CaCl2) so the bacteria can become competent

Cell Bio Lab Test#2 (4 decks)

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