Neonates that are kept in 100% oxygen incubators for several days become blind when they are removed from the incubator, a condition referred to as retrolental fibroplasia. What is the explanation for the loss of sight?

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#Question id: 16127

#Unit 13. Methods in Biology

You are studying regulation of the yeast enzyme glutamine synthetase (GS), which is encoded by the GLN1 gene. You have isolated two mutants, designated  gln2– and gln3–, that give decreased GS activity. Mating of either gln2– or gln3– haploids to wild type produces heterozygous diploids that show normal amounts of GS expression. When you cross either a gln2– or gln3– haploid to a gln1– strain the resulting diploids show normal expression of GS.

The GLN1 gene shows a rather complex regulation in response to different amino acids. When either glutamate (glu) or glutamine (gln) is added to the medium, the amount of GS expression diminished and when both glutamate and glutamine are added to the medium GS expression is shut off completely. The effects of different mutants on the response to glu and gln are shown below.

Which of the models from best fits these experimental results? Complete the model for the regulation of GLN1 that includes the effects of glu and gln.      

TLS Online TPP Program

#Question id: 16128

#Unit 13. Methods in Biology

You are studying regulation of the yeast enzyme glutamine synthetase (GS), which is encoded by the GLN1 gene. You have isolated two mutants, designated gln2– and gln3–, that give decreased GS activity. Mating of either gln2– or gln3– haploids to wild type produces heterozygous diploids that show normal amounts of GS expression. When you cross either a gln2– or gln3– haploid to a gln1– strain the resulting diploids show normal expression of GS. 

Next, you decide to evaluate the promoter for the GLN1 gene. To do this you first fuse the promoter region to the LacZ coding sequence and then place this hybrid gene on an appropriate yeast plasmid. You find that cells carrying the hybrid gene express activity under the same conditions that GS is expressed in wild type cells, meaning that the promoter region you have selected contains all of the necessary cis-acting sequences for normal regulation. The figure below shows the effect of different 50 bp deletions in the promoter region on the amount of ß-galactosidase activity expressed by the reporter gene. how would you expect a gln2– gln3– double mutant to behave?

TLS Online TPP Program

#Question id: 16129

#Unit 13. Methods in Biology

5 You are studying regulation of the yeast enzyme glutamine synthetase (GS), which is encoded by the GLN1 gene. You have isolated two mutants, designated gln2– and gln3–, that give decreased GS activity. Mating of either gln2– or gln3– haploids to wild type produces heterozygous diploids that show normal amounts of GS expression. When you cross either a gln2– or gln3– haploid to a gln1– strain the resulting diploids show normal expression of GS.

Describe the cis-acting elements in the GLN1 promoter that are evident from these experiments, giving both their position and as much of their function as you can deduce.

TLS Online TPP Program

#Question id: 16130

#Unit 13. Methods in Biology

You are studying regulation of the yeast enzyme glutamine synthetase (GS), which is encoded by the GLN1 gene. You have isolated two mutants, designated gln2– and gln3–, that give decreased GS activity. Mating of either gln2– or gln3– haploids to wild type produces heterozygous diploids that show normal amounts of GS expression. When you cross either a gln2– or gln3– haploid to a gln1– strain the resulting diploids show normal expression of GS.              

How many units of ß-galactiosidase would you expect to be expressed from deletion 1 in a gln2– mutant? 

TLS Online TPP Program

#Question id: 16131

#Unit 13. Methods in Biology

You are studying regulation of the yeast enzyme glutamine synthetase (GS), which is encoded by the GLN1 gene. You have isolated two mutants, designated gln2– and gln3–, that give decreased GS activity. Mating of either gln2– or gln3– haploids to wild type produces heterozygous diploids that show normal amounts of GS expression. When you cross either a gln2– or gln3– haploid to a gln1– strain the resulting diploids show normal expression of GS.              

 How many units of ß-galactiosidase would you expect to be expressed from deletion 4 in a gln2– mutant?

TLS Online TPP Program

#Question id: 16132

#Unit 13. Methods in Biology

You are running a human assisted reproduction clinic and providing state-of-the-art genetic diagnostic services. A married couple who already had a child with cystic fibrosis approach you because they wish to have another child, but only if they can be assured that the child will not have cystic fibrosis. You genotype the woman and discover that she is a heterozygote for Del508, the most common mutation causing cystic fibrosis. You suggest that the couple consider first polar body testing, in which several unfertilized oocytes (each with its first polar body) are retrieved from the woman, the first polar bodies are removed, and PCR tests are conducted on DNA from each of the first polar bodies. The couple agrees, and you obtain the following results:

 

the observation that the polar bodies for oocytes #1 and #5 test positive for both Del508 and the wild type sequence, because