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T2 Biology > Bacterial Transformation > Flashcards

Bacterial Transformation Flashcards

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1
Q

how can foreign genes be introduced into organisms

A
  • through bacterial transformation
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2
Q

what is the purpose of a plasmid during transformation

A

a plasmid is a small, circular piece of double-stranded DNA

-plasmid DNA contains coding sequences (genes) which are expressed by the bacterium. Often, the gene carried in plasmids provide bacteria with genetic advantages, such as antibiotic resistance

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3
Q

role of the beta-galactosidase (LacZ) (color marker gene)

A
  • breaks down lactose into galactose and glucose
  • can also cleave the synthetic analog of lactose, X-GAl
  • creates a product that is blue when cleaved with XGal
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4
Q

role of beta lactamase (ampicillin resistant gene)

A
  • it destroys the activity of ampicillin by breaking down the beta lactic ring
  • thus, bacteria with beta lactamase gene can resist ampicillin
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5
Q

purpose of nutrient agar

A

to contain the bacteria

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6
Q

purpose of ampicillin

A

ampicillin is a member of the penicillin family in antibiotics

  • destroys the activity of bacteria by destroying its cell wall
  • ampicillin contains a chemical group called BETA LACTAM RING which blocks the process by which bacteria link molecules to create a cell wall
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7
Q

purpose of X-gal

A
  • synthetic analog of lactose that when cleaved by the LacZ gene has a blue product
  • allows for the identification of ampicillin-resistant bacteria
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8
Q

purpose of pBlu

A

-a plasmid with all the necessary criteria to use in the bacterial transformation lab

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9
Q

what is bacterial transformation

A

when a host organism takes in foreign DNA and expresses its gene
- the introduction of apiece of DNA, like a plasmid, into a bacterial cell

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10
Q

what does a plasmid contain

A
  • origin of replication
  • gene for antibiotic resistance (encodes beta lactamase)
  • color marker gene (beta galactosidase/lactZ)
  • polylinker
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11
Q

what is a recombinant plasmid

A

a genetically engineered plasmid (with a foreign gene inserted in it)

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12
Q

why do we lower the temperature in the transformation experiment

A

to slow down the movement of the phospholipid in the membrane of the bacteria to efficiently neutralize the charges

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13
Q

why do we use the heat shock

A

rapid change in temperature induces the formation of pores in the bacterial membrane so that the plasmid DNA can enter

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14
Q

what are competent cells

A

when bacteria cells are in a state in which they are able to take up DNA, they are referred to as competent

-transformation rarely occurs naturally, so by subjecting bacteria to certain conditions, we can enable many of them to take up DNA

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15
Q
  • what is insulin +relation to bacterial transformation

- diabetes?

A
  • a hormone that originates from the pancreas, regulates glucose in blood
  • we can create insulin from bacteria by purifying it
  • diabetes consists of type 1 and type 2. It is a disorder that deactivates the insulin
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16
Q

how is a plasmid genetically engineered to contain a gene of choice

A

-a plasmid has sequences that restriction enzymes correspond to, so restriction enzymes can manipulate the plasmid to create a gene of choice by ‘cutting’ there

17
Q

why do we use CACl2 (calcium chloride) in the bacterial transformation lab

A
  • Plasmid DNA is a hydrophilllic molecule which will normally pass through the plasma membrane of the bacterium in order to accomplish transformation, the bacteria cells must first become competent to make the plasmid DNA.
  • this is done by neutralizing the negative charges of the membrane, the DNA molecule with calcium chloride
18
Q

origin of replication?

A
  • a sequences of DNA is recognized by DNA replication enzymes
  • when a bacterium divides all of the plasmids contained within the cell are replicated, thus each daughter cell receives a copy of each plasmid
19
Q

Results of bacterial transformation (with plasmid)

A
  1. (LB) lawn of bacteria
  2. (LB +AMP) bacterial growth ( white colonies)
  3. (LP+AMP+X-GAL) Blue colonies (and white)
20
Q

Results of bacterial transformation (without plasmid)

A
  1. (LB) lawn of bacteria
  2. (LB +AMP) nothing
  3. (LP+AMP+X-GAL)nothing
21
Q

where do transcription and translation take place

A

transcription: nucleus
translation: cytoplasm

T2 Biology (7 decks)

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