Name_____________________

Biology 50
Molecular Biology and Genetics
Exam 4
December 15, 1994


1. Describe position effect variegation. What is the structural basis for this effect and how was it discovered?


2. The following classes and frequencies of ordered tetrads were obtained from the cross a+ b+ x a b in Neurospora.

______________________________________________________________________________
Spore pair
							Number of Asci
______________________________________________________________________________

1-2 		3-4 		5-6		7-8			

a+b+		a+b+		ab		ab		1766

a+b+		a b		a+b+		ab		220

a+b+		a b+		a+b		ab		14


3. The following spore arrangements were obtained in the indicated frequencies from ordered tetrads in a cross between a Neurospora straion com/val which exhibits a compact growth form and is unable to synthesize the amino acid valine, and a wildtype strain +/+.

__________________________________________________________________________________________
Spore pair 			         Ascus composition
__________________________________________________________________________________________

1-2		c v 		c +		c v 		+ v		c v

3-4		c v		c +		c +		c +		+ v

5-6		+ +		+ v		+ v		c v		c +

7-8		+ +		+ v		+ +		+ +		+ +

Number:		34		36		20		1		9


What can you conclude about the linkage and location of the genes with respect to each other and to the centromere?

4. The gene met14 in the yeast S. cerevisiae codes for and enzyme used in the synthesis of the amino acid methionine and is very closely linked to the centromere of chromosome XI. Suppose that met14 (m) is used in mapping a new mutation, b, whose location in the genome is unknown. Diploids are made by mating m B x M b, in which the uppercase symbols denote the wildtype alleles, and asci are examined. In this mating, what kinds of asci correspond to parental ditype (PD), nonparental ditype (NPD) and tetratype (TT)? What relative proportions of PD, NPD and TT asci would be expected in the following cases:

a) b closely linked with m?

b) b on a different chromosome from that of m but close to the centromere?

c) b on a different chromosome from that of m and far from the centromere?

5. A gene R codes for a protein that is a negative regulator of transcription of a gene S. Is gene S transcribed in an R- mutation? How does the situation differ if the product of R is a positive regulator of S transcription?

6. For each of the following genotypes, state whether ß-galactosidase is made and whether its synthesis is inducible or constitutive.

a) lac Z+ lacY-/ lacz- lacY+

b) lacOc lacz- lacY+/ lacZ+ lacy-

c) lacp- lacZ+/ lacOc lacz-

d) lacI+ lacp- lacZ+/ laci- lacZ+

e) lacI+ lacp- lacY+/ laci- lacy-

7. Let Oo represent an operator mutation causing the operator to bind reversibly with the normal repressor. Let Rs representa a regulatory gene mutation that causes the formation of an altered repressor which cannot react with the inducer even though it can bind with the normal operator. For each of the following, tell which enzymes will be produced constitutively and which by induction?

a) Rs O+ Z+Y+A-

b) R+ Oo Z+ Y+ A+

c) R+ Oo Z+Y+A-/ r- O+ z- y- A+

d) r- O+Z+Y+a-/ Rs o-z-y-A+

8. Is it necessary for a gene that codes for the repressor of a bacterial operon to be near the structural genes? Why or why not?

9. Consider a eukaryotic transcriptional activator protein that binds to an enhancer sequence and promotes transcription. What change in regulation would you expect from a duplication in which several copies of the enhancer were present instead of just one?

10. Is an attenuator a region of DNA that, like an operator, binds with a protein? Is RNA synthesis ever initiated at an attenuator?

11. Compare and contrast three structural motifs in eukaryotic transcriptional activator proteins. How do the structures form and how do they facilitate DNA-binding?

12. Draw the essential elements of a eukaryotic promoter. Compare and contrast to prokaryotic promoter regions.

13. In normal mouse cells the protooncogene (p-onc) resides at a certain chromosome site. The DNA in the region of the protooncogene can be broken into specific fragments be a restriction enzyme, as indicated here by the arrows:

____________________________________________________________
p-onc


In the course of an experiment, normal mouse cells growing in culture are transformed into tumor cells following infection with a retrovirus. You wish to determine if the malignant transformation is the result of viral integration near p-onc. DNA is extracted from both normal and transformed cells. The DNA from both sources is exposed to a restriction enzyme, which fragments the DNA and attacks the sites indicated previously by arrows. The DNA is separated according to molecular weight, denatured and exposed to 1) denatured radioactive p-onc DNA and 2) denatured radioactive viral DNA. Autoradiographs are then prepared. Interpret teh comparison of the four autoradiographs shown here:


Cell source of 			Normal	Normal	Tumor   Tumor
mouse DNA   
         			_______________________________
						
Molecular weight				______	_____

				______
				_______________________________

Sources of radiolabeled
DNA:				p-onc	  virus	  p-onc  virus


14. Suppose in the case of another virus, the following results were obtained in a situation exactly like that described above. What would you conclude?

Cell source of 			Normal	Normal	Tumor   Tumor
mouse DNA   
         			_______________________________
						
Molecular weight				______	_____

				______
				_______________________________

Sources of radiolabeled
DNA:				p-onc	  virus	  p-onc  virus


15. Progression through the cell cycle depends on the interaction of two types of regulatory proteins: kinases and cyclins. List the functions of each and describe how they interact with each other to cause cells to move through the cell cycle.

16. How were cell cycle checkpoints discovered? How do the checkpoints relate to the cell cycle kinases and cyclins?

17. How would you describe cancer at the cellular level?



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Last modified: 1997 Sept 22
Comments to: (Yvonne Strickland)