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STUDY OBJECTIVE 1: To de ne bacteria and bacterial
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Prokaryotes (bacteria) usually have a single circular chromosome of double-stranded DNA. A bacteriophage consists of a chromosome wrapped in a protein coat. Its chromosome can be DNA or RNA. Phenotypes of bacteria include colony morphology, nutritional requirements, and drug resistance. Phage phenotypes include plaque morphology and host range. Replica-plating is a rapid screening technique for assessing the phenotype of a bacterial clone. STUDY OBJECTIVE 2: To study life cycles and sexual
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into the host chromosome, an Hfr cell results that can pass its entire chromosome into an F cell. The F factor is the last region to cross into the F cell. In transduction, a phage protein coat containing some of the host chromosome passes to a new host bacterium. Again, recombination with this new chromosomal segment can take place. STUDY OBJECTIVE 3: To make use of the sexual
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We can map the phage chromosome by measuring recombination after a bacterium has been simultaneously infected by two strains of the virus carrying different alleles. In E. coli, mapping is most ef ciently accomplished via interrupted mating and transduction. The former provides information on general gene arrangement and the latter provides ner details.
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In transformation, a competent bacterium can take up relatively large pieces of DNA from the medium. This DNA can be incorporated into the bacterial chromosome. During the process of conjugation, the fertility factor, F, is passed from an F to an F cell. If the F factor integrates
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Tamarin: Principles of Genetics, Seventh Edition
II. Mendelism and the Chromosomal Theory
7. Linkage and Mapping in Prokaryotes and Bacterial Viruses
The McGraw Hill Companies, 2001
Seven
Linkage and Mapping in Prokaryotes and Bacterial Viruses
S O L V E D
PROBLEM 1: A wild-type strain of B. subtilis is transformed by DNA from a strain that cannot grow on galactose (gal ) and also needs biotin for growth (bio ). Transformants are isolated by exposing the transformed cells to minimal medium with penicillin, killing the wildtype cells. After the penicillin is removed, replica-plating is used to establish the genotypes of 30 transformants:
Class 1 gal Class 2 gal Class 3 gal bio bio bio 17 4 9
P R O B L E M S
Class 1 gal Class 2 gal Class 3 gal Class 4 gal bio bio bio bio att att att att 71 0 9 26
What is the gene order, and what are the relative cotransduction frequencies Answer: We have selected all transductants that are gal . Class 2 is in the lowest frequency (0) and therefore represents the quadruple crossover between the transducing DNA and the host chromosome. From this, we see that att must be in the middle because this lowprobability event is the one that would have switched only the middle locus. In other words, the two end loci would be recombinant, and the middle locus would have the host allele. We can only calculate two cotransduction frequencies because these are selected data. Note that in class 1, there is no cotransduction between gal and either of the other two loci; class 2 would show the cotransduction of gal and bio; class 3 represents the cotransduction of gal and att ; and class 4 represents the cotransduction of gal and both other loci. Therefore, cotransduction values are gal att gal bio (9 (0 26)/106 26)/106 35/106 26/106 0.33 0.25.
What is the relative co-occurrence of these two loci Answer: The three classes of colonies represent the three possible transformant groups. Classes 2 and 3 are single transformants and class 1 is the double transformant. We are interested in the relative co-occurrence of the two loci. Therefore we divide the number of double transformants by the total: r 17/(17 4 9) 0.57. This is a relative value inverse to a map distance; the larger it is, the closer the loci are to each other.
PROBLEM 2: A gal bio att strain of E. coli is transduced by P22 phages from a wild-type strain. Transductants are selected for by growing the cells with galactose as the sole energy source. Replica-plating and testing for lysogenic ability gives the genotypes of 106 transformants:
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