Exp 3 Flashcards

(31 cards)

1
Q

what is an operon

A

it is a group of genes that are involved in a given function and are regulated by a single promoter

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2
Q

for genes under an operon, how are they transcribed

A

they are regulated together and are transcribed into a single mRNA transcript that is translated into different protein products

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3
Q

what does an operon consist of

A

more than one gene
a single promoter
regulatory binding sites

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4
Q

in a lac operon what is the role of regulatory binding sites

A

they are regions in the DNA where regulatory proteins bind and influence RNA polymerase

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5
Q

where are operons found? in what organism

A

they are found in prokaryotes only

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6
Q

why are operons not found in eukaryotes

A

eukaryotic genes carry their own individual promoters and are regulated independently

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7
Q

what is the lac operon

A

this operon contains genes that are involved in lactose metabolism

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8
Q

what is the purpose of lactose metabolism

A

it allows lactose to be used as an energy source to derive glucose when glucose is absent

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9
Q

why is the lac operon not always active

A

cells prefer to use all available glucose and only use lactose when nevessaru

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10
Q

what is beta-galactosidase for?

A

it is an enzyme that cleaves the disaccharide lactose into glucose and galactose which can be further converted into glucose

yield = 2 glucose molecules

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11
Q

what is the cap site and operator

A

these are the regulatory binding sites

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12
Q

what is the role of the cap site

A

it is where the CAP binding protein binds in the absence of glucose

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13
Q

what is the role of the operator

A

it is where the repressor comes OFF in the presence of lactose

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14
Q

what is the bond between galactose and glucose in lactose

A

a beta bond

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15
Q

what is the purpose of beta bond cleavage

A

to produce glucose molecules

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16
Q

what is the car brakes and accelerator analogy with the lac operon

A

the car can only go when the brakes are let off (the supressor) and when the gas pedal is on (the CAP binds)

17
Q

when will transcription occur in the lac operon

A

when the CAP binds in the absence of glucose

when the Suppressor releases in the presence of lactose

18
Q

do the CAP and operator have any affect on eachother

A

NO they are regulated independently

19
Q

what is the purpose of sucrose in this experiment

A

the lac operon is specific to lactose only

it will not produce an enzyme that cleaves sucrose

20
Q

if the glucose is present and lactose is not, what are the states of the activator (CAP) and repressor?

A

CAP is off

Repressor is on

21
Q

if the glucose and lactose are present, what are the states of the activator (CAP) and repressor?

A

CAP is off
repressor is off

22
Q

if no glucose or lactose is present, what are the states of the activator (CAP) and repressor?

A

CAP is on
repressor is on

23
Q

if no glucose is present but lactose is , what are the states of the activator (CAP) and repressor?

A

CAP is on
Repressor is off

24
Q

For the media containing glucose, do we expect high cell growth? what about activation of the Lac Operon and enzyme expression?

A

high cell growth, no activation

25
For the media containing sucrose, do we expect high cell growth? what about activation of the Lac Operon and enzyme expression?
no cell growth, no activation
26
For the media containing lactose, do we expect high cell growth? what about activation of the Lac Operon and enzyme expression?
high cell growth, activation
27
For the media containing water do we expect high cell growth? what about activation of the Lac Operon and enzyme expression?
no cell growth, no activation
28
what is the role of ONPG
it has a similar bond to lactose and will be cleaved to form ONP which has a yellow color that can be measured to analyze enzyme activity
29
why cant we use regular catastrophic reagents
they will denature the proteins
30
why do we use toulene
it pokes holes in the cells and allows the beta-galactosidase to leak out
31
what is measured by a spectrophotometer
the amount of light absorbed, it allows us to measure the amount of ONP and cell density