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Lac Operon Lab Answer

lac operon lab

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0% found this document useful (0 votes)
35 views10 pages

Lac Operon Lab Answer

lac operon lab

Uploaded by

meghna patel
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
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Question 3

According to Storhaug et al. (2017), approximately what percentage of


humans are lactase persistent?
Ans: 32
Question 4
Match the term from the drop-down menu with its best description.
Sugar in mammalian milk
Ans: Lactose
Enzyme in humans that breaks down the sugar in milk
Ans: Lactase
Enzyme in Escherichia coli bacteria that breaks down the sugar in milk
Ans: beta-galactosidase
Indicator added to student assay to determine concentration of bacterial
enzyme
Ans: ONPG
Turns yellow in aqueous solution
Ans: ONP
Chemical added to experimental tubes to kill the bacteria
Ans: Toluene
One of the 2 monomers that make up the sugar in milk
Ans: Galactose
Gene in humans that produces the enzyme that breaks down the sugar in
milk
Ans: LCT
Gene in Escherichia coli bacteria that produces the enzyme that breaks
down the sugar in milk
Ans: lacZ
Gene in humans that regulates production of the enzyme that breaks down
milk sugar
Ans: MCM6
Gene in Escherichia coli that was tested to determine if it regulates
production of the enzyme that breaks dow milk sugar
Ans: lacI
Question 5
How many mutations are known to cause lactase persistence in humans?
Ans:5
Question 6
In humans, intron 9 of the [ MCM6] gene, which is an
[unexpressed] region of the gene, contains a mutation that causes
lactase persistence. This region is [ an enhancer]
of the [ LCT] gene; therefore, it attracts
[ activator proteins] that increase production of the enzyme,
[ LCT]
Question 7
In this experiment, we are testing 2 things:
1. Can [ a bacterial species] turn a gene called [ lacZ] on and off,
and therefore sometimes produce [ beta-galactosidase] and
sometimes not; and
2. Is the [ lacI] gene [ an enhancer] in [Eschericha coli] like
[ MCM6] is in [ humans]
Question 8
The lac operon comprises (mark all that apply)
Group of answer choices
Ans: all the following are part of lac operon
lacZ.
a promoter.
the gene for beta-galactosidase.
an operator.
genes for 3 different proteins.
Exclude : ENHANCER, the gene for the lac repressor protein and lac I *
Question 9
The lac repressor protein is produced by
Group of answer choices
Ans: the lacI gene
Question 10
Consult the experimental details in the lab protocol. Then match the
contents listed with the proper tube from the drop-down menu.
Group of answer choices
1
[M9 media, wildtype E. coli, water]

2
[M9 media, wildtype E. coli, lactose]

3
[ M9 media, wildtype E. coli, glucose]

4
[M9 media, lacZ negative E. coli, water]

5
[M9 media, lacZ negative E. coli, lactose]

6
[M9 media, lacZ negative E. coli, glucose]

7
[M9 media, lacI negative E. coli, water]

8
[ M9 media, lacI negative E. coli, lactose]

9
[M9 media, lacI negative E. coli, glucose]

10
[M9 media, water]

Question 11
Which of the graphs above would be the best match for the wild-type
strain's response if E. coli turns on the lacZ gene in the presence of sugars
but not water?
Group of answer choices
Ans: D
Question 12
Which of the graphs above would be the best match for the lacZ-- strain's
response if E. coli turns on the lacZ gene in the presence of sugars but not
water?
Group of answer choices
Ans: None of these; the lacZ-negative strain should not produce beta-
galactosidase.

Question 13
Which of the graphs above would be the best match for the lacI-- strain's
response if the lacI gene were an enhancer for lacZ like MCM6 is for LCT?
Ans:G
Question 14
Suppose, for the sake of argument, that E. coli turned on the LacZ gene
only in the presence of lactose and nothing else. Which of the graphs
above would best support that conclusion in the wild-type strain?
Group of answer choices
Ans: B

Question 15
Consider graph (G) above. Suppose that was the result observed for the
lacI-- strain. Which of the following would be a proper conclusion that
could be made from that observation?
Ans:The lacI gene is precisely the opposite of an enhancer; it is an
inhibitor.
Question 16
Consider graph (F) above. Suppose that was the result obtained for the
lacZ-- strain. Which of the following would be a proper conclusion that
could be made from that observation?
Ans: The experiment is seriously flawed, so the results are unreliable.
Question 17
Compare your data to the predictions in the previous questions. Do your
data support, contradict or say nothing about the hypothesis that E.\ coli
upregulates (turns on) the lacZ gene in the presence of sugars but not
otherwise? Explain your reasoning fully and explain how the data apply to
your argument. Be as specific and complete as you can.
Ans: yes, our data support the prediction that ‘E. coli upregulates (turns
on) the lacZ gene in the presence of only lactose sugar but not otherwise’.
It can be said so as in tube no. 2, which has wild type culture of E.coli with
lactose sugar showed high level of enzyme activity, as compared to tube 1
which has no sugar and tube 3 which has glucose as a sugar but not
lactose.
Question 18
Again, compare your data to your predictions. Do your data support,
contradict or say nothing about the hypothesis that the lacI gene is an
enhancer of lacZ in E. coli like MCM6 is an enhancer of LCT in humans?
Explain your reasoning fully and explain how the data apply to your
argument. Be as specific and complete as you can.

Ans: Our data contradict the hypothesis that lacI gene is an enhancer of
lacZ in E. coli unlike MCM6 is an enhancer of LCT in humans.
It can be said so, as if the bacteria have normal lac I gene, it prevents the
constitutive expression of lac Z gene which results in very low levels of
enzyme, which can be concluded by comparing the results of 2 strains of
E.coli, i.e. wild type and lac I-. Mutation in lac I gene cause continuous
production of the beta galactosidase enzyme in all the cases, regardless
presence or absence of any specific sugar. So it can be said that in normal
condition lac I gene product act as repressor and not enhancer.
However, if we compare the data generated by MCM mutant and the lac I
mutant they look similar, but the difference is that mutation in MCM has
increased its action but mutation in lac I results in loss of its action. So no
lac repressor produced, so no binding of repressor to the promoter of lac
operon, so the promoter is free, so the enzyme RNA polymerase can bind
the promoter and transcribe the mRNA of the beta galactosidase enzyme
which increases the level of the enzyme after translation step.

Question 19
In your textbook, study the section on negative regulation of the lac
operon (section 18.3).
In the space provided, answer the following question:
Do your data support, contradict or say nothing about the lac operon
hypothesis presented in your text? Explain your reasoning fully and
explain how the data apply to your argument. Be as specific and complete
as you can.
Ans: As per lac operon hypothesis in the text “if lac Z and lac Y genes are
under the negative control” and “ does lac I produces a product that
represses the transcription of lac Z and lac Y gene’.
Our data supported both the hypothesis.
Lac Z is the gene for synthesis of beta galactosidase which has been
estimated in this experiment. So if the gene is functional it produces the
enzyme, if it is repressed it cannot produce the enzyme.
In wild type culture of E.coli, the enzyme level was high when the medium
had lactose (tube no 2), which shows that lactose should be the anti-
repressor molecule which binds to repressor and allow the transcription of
the enzyme’s mRNA. In other cases the level of the enzymes were low
(tube 1 and tube 3 with wild type E.coli), which shows that the lacZ gene
is under negative control. However, no data have been collected for lac Y
action.
The results of lac I- strain of E.coli shows high level of the enzymes,
which are indicative of the fact that in absence of lac I gene product, the
operon will be functional to produce more mRNA and more protein, here
beta galctosidase enzyme which is observed in tube no 7,8 and 9 as
compare to the wild type strain of E.coli. So it can be said that lac I
produces a product that represses the transcription of lac Z and lac Y
gene’.

Question 20
Why did the students add toluene and sarkosyl detergent to the tubes?
Ans: To kill the bacteria and release the enzyme into solution.
Question 21
The standardized amount of beta-galactosidase per bacteria in tube A is
1.45
Question 22
The standardized amount of beta-galactosidase per bacteria in tube B is
2.36
Question 23
These data support the following conclusions:
 Tube A has [more] bacteria compared to
tube B.
 Tube A has [more] beta-galactosidase
compared to tube B.
 Bacteria in tube A are producing beta-galactosidase
[ at a lower rate than] bacteria in tube B.

Question 24
In the space provided, explain your answer to the previous question. What
reasoning led you to choose the answers you did?

Ans: The greater absorbance of tube A (0.95) after incubation of mixture


containing M9 media, lactose and E.coli bacteria as compared to tube B
(0.22) shows that the number of bacteria present in tube A is higher than
tube B.
Here the absorbance is due to the density of the mixture, which is due to
presence of bacteria.

At 420 nm, after adding the lysing reagent and indicator ONPG, higher
absorbance in tube A (1.38) as compared to tube B (0.52), shows that
more production of ONP in tube A which is yellow colored. More ONP
would have been formed due to higher levels of beta galactocidase enzyme
which is the catalyst of the reaction. Thus over all higher level of the
enzyme in tube A.

If we calculate the standardized amount of beta-galactosidase produced per


bacteria by the formula of absorbance at 420 nm (which is due to presence
of ONP) / absorbance of the tube before addition of the indicator ( which is
due to the presence of bacteria), the value we get for tube A is 1.45 and
tube B is 2.36. Thus even though the total value of the enzyme present in
tube A is greater than tube B, the rate at which the enzyme produced in
tube A is slower than that of tube B.

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