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Tamarin: Principles of Genetics, Seventh Edition
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III. Molecular Genetics
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12. DNA: Its Mutation, Repair, and Recombination
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The McGraw Hill Companies, 2001
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DNA: Its Mutation, Repair, and Recombination
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a. Draw a deletion map of these mutations. b. A point mutation, 6, is isolated and crossed with all of the deletion strains. Wild-type recombinants are recovered only with strains 2 and 3. What is the location of the point mutation 20. Hydroxylamine is a chemical that causes exclusively C T transition mutations. Can hydroxylamine reverse nonsense mutations Explain. 21. A nonsense suppressor is isolated and shown to involve a tyrosine transfer RNA. When this mutant transfer RNA is sequenced, the anticodon turns out to be normal, but a mutation is found in the dihydrouridine loop. What does this nding suggest about how a transfer RNA interacts with the messenger RNA 22. Devise selection-enrichment procedures for isolating the following kinds of mutants: a. extra-large bacterial cells b. nonmotile ciliated protozoans 23. Two chemically induced mutants, x and y, are treated with the following mutagens to see if revertants can be produced: 2-amino purine (2AP), 5-bromouracil (5BU), acridine dye (AC), hydroxylamine (HA), and ethylmethanesulfonate (EMS). In the following table, revertants and no revertants. For each mutation, determine the probable base change that occurred to change the wildtype to the mutant.
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Chemical Mutant x y 2AP 5BU AC HA EMS
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25. What situation will lead to false negatives in a complementation test or, in other words, indicate mutations are in the same gene when, in fact, they are in different genes 26. Suppose you repeat the Luria-Delbr ck uctuation test, but this time you look for lac colonies. Your individual cultures produce the following numbers of lac colonies: 20, 25, 22, 18, 24, 19, 17, 25, 26, and 18. Subsamples from the bulk culture give results identical to these. What can you conclude 27. You have isolated a new histidine auxotroph, and, despite all efforts, you cannot produce any revertants. What probably happened to produce the original mutant
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DNA REPAIR
28. UV light causes thymine dimerization. Describe the mechanisms, in order of ef ciency, that can repair the damage. Name the enzymes involved. 29. What types of damage do excision repair endonucleases recognize 30. What are the functions of the RecA protein How is it involved in phage induction (See also RECOMBINATION)
RECOMBINATION
31. Diagram, in careful detail, a recombination by way of the double-strand break model.What enzymes are required at each step 32. What are the different enzymes involved in reciprocal and nonreciprocal recombination
24. What situation will lead to a false positive in a complementation test or, in other words, indicate two genes when, in fact, the mutations are in the same gene
C R I T I C A L
T H I N K I N G
Q U E S T I O N S
1. Charles Yanofsky demonstrated colinearity of the gene and its protein product. What are the alternatives to colinearity
2. Comment on the statement that DNA is a molecule designed for replication and repair.
Suggested Readings for chapter 12 are on page B-10.
Tamarin: Principles of Genetics, Seventh Edition
III. Molecular Genetics
13. Genomics, Biotechnology, and Recombinant DNA
The McGraw Hill Companies, 2001
GENOMICS, BIOTECHNOLO GY, AND RECOMBINANT DNA
STUDY OBJECTIVES
1. To look at the techniques of gene cloning 358 377 2. To examine the techniques of creating restriction maps 3. To study the methods of DNA sequencing 383 4. To look at the goals and methods of the Human Genome Project 390 5. To look at the practical bene ts and human issues of genetic engineering 397
STUDY OUTLINE
Genomic Tools 359 Restriction Endonucleases 359 Prokaryotic Vectors 360 Cloning a Particular Gene 364 Southern Blotting 366 Probing for a Cloned Gene 368 Eukaryotic Vectors 369 Expression of Foreign DNA in Eukaryotic Cells 372 Restriction Mapping 377 Constructing a Restriction Map 379 Double Digests 379 Restriction Fragment Length Polymorphisms 380 Polymerase Chain Reaction 381 DNA Sequencing 383 The Dideoxy Method 383 Creating a General-Purpose Primer 386 Mapping and Sequencing the Human Genome 390 Locating a Gene of Interest 390 The Human Genome Project 391 Ethics 397 Practical Bene ts from Gene Cloning 397 Medicine 397 Agriculture 398 Industry 398 Summary 398 Solved Problems 399 Exercises and Problems 400 Critical Thinking Questions 404 Box 13.1 The Recombinant DNA Dispute 370 Box 13.2 Cloning Dolly 374 Box 13.3 Genes Within Genes 388
Arti cially colored transmission electron micrograph of DNA plasmids from the bacterium Escherichia coli. These plasmids are used in genetic engineering.
( Dr. Gopal Murti/SPL/Photo Rsearchers, Inc.)
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