lab 9 Flashcards

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1
Q

what does genetic transformation mean

A

change caused by genes

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2
Q

what gene are we looking at

A

GFP - Green Fluorescent Protein

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3
Q

real-life source of GFP

A

jellyfish Aequorea victoria

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4
Q

Bio-Rad’s pGLO plasmid encodes for

A

gene GFP and gene for resistance to antibiotic ampicillin

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5
Q

what sugar are we using

A

arabinose

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6
Q

what effect does arabinose have on the expression of GFP

A

it turns it on

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7
Q

transformed cells will appear _________ on plates not containing arabinose

A

white

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8
Q

transformed cells will turn ___________ under UV light when arabinose is included

A

fluorescent green

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9
Q

arabinose is a source of of what for bacteria

A

energy and carbon

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10
Q

how does arabinose initiate transcription of the genes to break it down

A

promotes binding of RNA polymerase

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11
Q

in the genetically engineered pGLO plasmid, some of of the genes involved in the breakdown of arabinose are replaced with what?

A

GFP

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12
Q

3 genes that code for digestive enzymes

A

araB, araA, araD

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13
Q

what are araB, araA, araD clustered in

A

arabinose operon 3

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14
Q

promoter of araB, araA, araD

A

PBAD

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15
Q

araC protein

A

binds to DNA at binding site for RNA pol

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16
Q

what is present in the DNA code of the pGLO plasmid?

A

promoter (PBAD) and araC

17
Q

what does aran promote the binding of

A

RNA polymerase

18
Q

LB

A

liquid/solid nutrient media

19
Q

what is LB named after

A

Luria and Bertani

20
Q

what is agar derived from

A

seaweed

21
Q

beta-lactamase gene

A

provides resistance to antibiotic ampicillin

22
Q

Ca2+ of transformation solution purpose

A

neutralizes negative charges of phosphate backbone and phospholipids of cell membrane to allow DNA to enter the cells

23
Q

what does heat shock do

A

increases permeability of membrane to DNA

24
Q

what times are we looking at the plates

A
  • 18-24
  • 24 hours before next lab
25
Q

transformation efficicency

A

ug of DNA spread on plate