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1.The function of six hypothetical genes in a microbial organism are shown below

ID: 18325 • Letter: 1

Question

1.The function of six hypothetical genes in a microbial organism are shown below:

Gene    Function of the type of gene

GRA     Required for the utilization of the sugar granulose as a carbon source

RAC     Required for sensitivity to the drug ractocine

IOB      Required for the synthesis of the vitamin iobine

YYZ      Required for resistance to YYZ virus   

PET    Required for the synthesis of the growth hormone petozone

AOA Required for the growth of hyphae from the cell            

       Based on the function of these genes suggest one possible genetic screen to identify a loss-­of-­function mutation in each of these genes.

2.       You construct the following genetic map distances between these genes

           Gene GRA RAC    IOB     YYZ     PET    AOA

GRA    _    2.5 0.1     1.9     0.7 2.0

RAC    2.5 _    2.6    0.6    3.2 0.5

IOB     0.1 2.6    _    2.0    0.6 2.1

            YYZ       1.9       0.6     2.0       _      2.6     0.1

            PET         0.7       3.2   0.6      2.6     _       2.7

            AOA      2.0       0.5     2.1      0.1     2.7   _

Base on these map distances construct a genetic map that includes all of these genes.

3. You decide to revert the iob-­ mutant to wild type (IOB+) using different mutagens and find the following reversion rates for each mutagen tested

Mutagen                 Rate of reversion to IOB+ (%)

2-­aminopurine                 98
5-­bromouracil                  95
Nitrous acid                      94
Proflavin                             2
Spontaneous                       4

What is the most likely molecular basis of the original iob-­ mutation?


What does this reversion test data suggest about the most prominent mechanism of spontaneous mutations in this organism?

4.You decide to isolate deletion mutations in the IOB gene. Suggest a genetic screen that would enrich for deletion mutations in this gene over other types of loss-­of-­function mutations.

5. Suggest two genetic tests that would show you have indeed identified iob deletion mutants?

6. It is generally understood that deletion a gene result in a null phenotype for that gene function. Assuming you have a deletion of IOB gene you now want to determine if the original iob loss-­of-­function mutant you identified in Question 1 is a true null mutant. How can you use the deletion mutant you found in Question 4 to do this?

Explanation / Answer

1.) GRA: put organisms in a medium with sugar granulose as the only carbon source RAC: IOB: test for the organisms with iobine YYZ: introduce YYZ virus PET: test for growth hormone AOA: test for hyphae

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