Tutorial 1 Q: A species of cereal rye (Secale cereale) has a chromosome number of 14, while a species of Canadian wild rye

(Elymus canadensis) has a chromosome number of 28. Sterile hybrids can be produced by crossing Secale with Elymus. a. What would be the expected chromosome number in the somatic cells of the hybrids? b. Assume that the G1 nuclear DNA content of Elymus is 25.5 picograms and that the G1 nuclear DNA content of Secale is 16.8 picograms. What would be the expected DNA content in a metaphase somatic cell of the hybrid? c. Why are these hybrids sterile? Answer: a. Cereal rye produces gametes with a chromosome number of 7, and Canadian wild rye produces gametes with a chromosome number of 14. When these two gametes fertilized, the expected chromosome number in the somatic cells of the hybrids are 7+14=21. During G1 phase, the number of chromosomes have not duplicated yet. If the DNA content of Elymus is 25.5, so the DNA content for the germ cells is 25.5/2=12.75 and for Secale, it is 16.8/2=8.4. Therefore, the DNA content in the hybrid of these two species would be 12.75+8.4 = 21.15. Since at metaphase, the number of chromosomes has doubles, so the expected DNA content will be 21.15x2=42.3 picograms. Because meiosis is arrested in prophase I due to the failure of pairing of homologous chromosomes, no gametes will be produced.

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Tutorial 2 Could nad-110 repair nadB164? ANSWER: No. Deletion removes interval containing wild-type equivalent of allele 164. Could nad-114 repair nadB95? ANSWER: Yes. Deletion does not remove wild-type equivalent of allele 95. Could nad-110 repair nad-114? ANSWER: No. Region corresponding to 110 would still be missing. Could nadB172 repair nadB95? ANSWER: Yes. Each point mutant contains the wild-type interval for the other allele. Could nadB172 repair nadB62? ANSWER: Maybe. Even though both alleles map within the same deltion interval, you do not know whether the mutations in alleles nadB172 and nadB62 affect the same base pair or not. If the mutations

affect the same base pair, recombinational repair would not be possible. However, if the mutations affect adjacent base pairs or base pairs that are further apart, recombinational repair is possible -- the frequency of recombinational repair would depend upon how far apart the two mutations are, so if the two mutations are very close this would be a rare event. Why are there so many mutations in some deletion intervals and not in others? ANSWER: Different parts of a gene may are susceptible to a mutagen differently. In other words, each mutagen will may "hot spots" for mutagenesis which may not be evenly distributed throughout the gene. Not all residues of the gene product are essential for proper structure and/or equally important for its function. The codons corresponding to the critical residues will be the only mutable sites recoverable as missense mutants. Such critical residues are often unevenly distributed within the gene.

You isolate a new mutant that had a phenotype expected for a nadB mutation but could repair all of the known nadB deletion mutations. Suggest two potential explanations for this result? ANSWER: The mutant may be affected in a gene other than nadB but which confers a NadB- phenotype when mutated. Other genes in a common pathway or trans-acting regulatory factors for nadB might have this phenotype. In this case the nadB gene in the donor would be wild-type and, hence, would be able to repair all deletion intervals of nadB in the recipient by recombinational repair. The new mutant allele may be located within the nadB gene but maps outside of the end points of the deletion intervals, for example in the same interval as allele 55.

What will be the constituents of the medium used to select exconjugants that have acquired the thr+ allele? Medium must contain ampicillin to eliminate donors which are sensitive to this antibiotic. Medium must also contains glucose and all amino acids for the genes being mapped, except threonine, which will allow only exconjugants having the thr+ allele transferred from the donor to grow, while exconjugants without Thr+ and the initial recipients (F- strain) will not grow.

What is the order of the three markers? Explain why. The order of the gene is Arg-His-Leu OR Leu-His-Arg. The data show that His has much higher cotransduction rate with Arg, so His is closer to Arg than Leu. What are the cotransduction frequencies? Cotransduction frequency is the percentage of cells that received two markers.

For arg and his, this includes the Arg+ Leu His+(307) and Arg+ Leu+ His+(112). The cotransduction frequency of arg and his is 419 (2nd gene)/1000 (1st gene), equals 41.9% As the same, cotransduction frequency of arg and leu is 113 (2nd gene)/1000 (1st gene), equals 11.3%