You have isolated 8 mutants in yeast that fail to grow on minimal media plates but do grow when they are supplemented with Arginine. You know that Arginine is synthesized in a biochemical pathway within wild-type yeast, but you do not know how many gene products it takes for the pathway. You have all of the lines as both a and a cells and mate each strain to each other in pairwise crosses and plate them on minimal media to see if they grow. You obtain the following results with (+) representing growth, and (-) indicating no growth: a 1 5 1 3 4 7 How many genes are represented? 2 O 3 4 O 5 8 9 Cannot tell from the data a + + + + + . + +, 0 +, + + + . + + - + + + + + i++ ni + + + + + . + + + m + + + + . + + + N +, + + + , + + + . + + +
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- When 1 million cells of a culture of haploid yeastcarrying a met− auxotrophic mutation were plated onpetri plates lacking methionine (Met), five coloniesgrew. You would expect cells in which the originalmet− mutation was reversed (by a base change back tothe original sequence) would grow on the media lackingmethionine, but some of these apparent reversions couldbe due to a mutation in a different gene that somehowsuppresses the original met− mutations. How wouldyou be able to determine if the mutations in your fivecolonies were due either to a precise reversion of theoriginal met− mutation or to the generation of a suppressor mutation in a gene on another chromosome?The completely synthetic yeast chromosome Syn IIIcontains a loxP site in the 3′ UTR of every gene thatis potentially nonessential to yeast survival. As youwill recall from Chapter 6, loxP sites are targets ofsite-specific recombination. The researchers who constructed Syn III included these loxP sites as a way to“scramble” the chromosome, meaning that parts ofthe chromosome could easily be deleted or rearranged.The goal of these investigations is to drive the evolution of Syn III so as to define a minimal genome thatcan support the life of this organism. Outline the experiment the researchers would do to scramble Syn IIIin order to define a minimal genome.By conducting conjugation experiments between Hfr and recipientstrains, Wollman and Jacob mapped the order of many bacterialgenes. Throughout the course of their studies, they identified severaldifferent Hfr strains in which the F-factor DNA had been integratedat different places along the bacterial chromosome. A sample of theirexperimental results is shown in the table:Draw a map that shows the order of genes and the locations ofthe origins of transfer among these different Hfr strains?
- In E. coli, four Hfr strains donate the following markers,shown in the order donated:Strain 1: M Z X W CStrain 2: L A N C WStrain 3: A L B R UStrain 4: Z M U R BAll these Hfr strains are derived from the same F+ strain.What is the order of these markers on the circularchromosome of the original F+?The following recombinants are recovered when conjugation occurs between an a*d*g+ donor and an adg recipient. at d+ g+ = 84% a d g+ = 6% at d g+ = 10% a dt g+ = less than 1% What is the map distance between the a and d genes? 10 map units 74 map units less than 1 map unit 84 map units 6 map unitsAn Hfrstrain that is a *b*c*d* e*f* g *h* is mated with an F strain that is a b e d e f gh. The mating is interrupted at 5 minutes interval, and the genotypes of the F recombinants are determined. The results obtained are tabulated in Table 2. Draw the map of the Hfrchromosome and indicate the position of the origin of transfer, the direction of the transfer and the minutes between genes. Table 2:Entry time of Hfr chromosome into recipient cell. Time a d e f h 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + +
- Streptomycin resistance in Chlamydomonas may result from a mutation in either a chloroplast gene or a nuclear gene. What phenotypic results would occur in a cross between a member of an mt+ strain resistant in both genes and a member of a strain sensitive to the antibiotic? What results would occur in the reciprocal cross?By conducting conjugation experiments between Hfr and recipientstrains, Wollman and Jacob mapped the order of many bacterialgenes. Throughout the course of their studies, they identified severaldifferent Hfr strains in which the F-factor DNA had been integratedat different places along the bacterial chromosome. A sample of theirexperimental results is shown in the following table:Analyze data. Compare and contrast. Make a drawing.You have a strain of Neurospora that is unable to synthesize histidine and thus requires H in the media in order to grow. You have isolated one revertant colony. Predict the expected proportion of the progeny that would be h+ if you cross the colony with the original mutant colony and the reversion occurred by each of the following mechanisms: Precise change of the mutated base back to its original base. A suppressor gene is mutated on a different chromosome A suppressor gene is mutated on the same chromosome but 10mu distant from the mutated gene. The mutant colony is crossed to a wild-type Neurospora colony and the following data are collected. 95% of all asci scored are h+ but 5% are h-. Which mechanism in part a is consistent with these data? Explain why and what has happened on a molecular level.
- Schizosaccharomyces pombe, also known as "fission yeast," is a powerful model organism in molecular and cell biology. While performing a genetic screen, you discover an auxotrophic S. pombe strain that is unable to synthesize one or more vitamins. The following table represents the key experiments you performed during your genetic screen. Fill in the table with the outcome of each experiment for your mutant strain (using + for growth and - for no growth). Medium Rich media Minimal media Minimal media + all vitamins Minimal media + all amino acids Growth Wild-type + + + + Mutant + + + > > >A prototrophic strain (his" arg" lac') was used as a donor to transform an auxotrophic strain (his arg lac'). Initial transformants are isolated on minimal medium + histidine + arginine + lactose - glucose. i. What genotypes will grow on this medium? ii. These colonies are replicated to minimal medium + histidine, and 50% of the original colonies grow. What genotypes will grow on this medium? iii. The original colonies are also replicated to minimal medium + arginine, and 10% of the colonies grow. What genotypes will grow on this medium? iv. The original colonies are also replicated to minimal medium. No colonies grow. Based on this information, what genotypes will grow on minimal medium + histidine and on minimal medium + arginine? What is the relative gene order for his, arg, and lac? Which two genes are closer? Explain your answer.An Hfr strain that is leuA+ and thiL+ was mixed with a strain thatis leuA− and thiL−. In the data points shown in the following graph,the conjugation was interrupted at different time points, and thepercentage of recombinants for each gene was determined bystreaking on a medium that lacked either leucine or thiamine.What is the map distance (in minutes) between these two genes?