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Which of the following two maps is consistent with the results
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Tamarin: Principles of Genetics, Seventh Edition
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III. Molecular Genetics
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13. Genomics, Biotechnology, and Recombinant DNA
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The McGraw Hill Companies, 2001
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Exercises and Problems
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30. You now take an unlabeled molecule from problem 29, digest it with HindIII, and get two fragments, 5.5 and 2.5 kb in size. If HindIII does not cut within the 3.5 kb EcoRI fragment, what size fragments do you expect in a double digest of HindIII and EcoRI 31. Two normal individuals have a child with Down syndrome. RFLP analysis with a probe from chromosome 21 is performed on all three individuals, and the results of the gels appear as follows. Based on these results, what can you conclude about the origins of the number 21 chromosomes
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34. The following diagram is of a dideoxy sequencing gel. What is the sequence of the DNA under study
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32. What is PCR When is it used
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DNA SEQUENCING
33. What are the steps in the dideoxy method of DNA sequencing How has the technique been improved with uorescent dyes
35. How can a particular piece of DNA be manipulated to be in the appropriate con guration for dideoxy sequencing 36. Provide, if possible, DNA sequences that can mark the termination of one gene and the initiation of another, given that the genes overlap in one, two, three, four, ve, six, or seven bases.
Tamarin: Principles of Genetics, Seventh Edition
III. Molecular Genetics
13. Genomics, Biotechnology, and Recombinant DNA
The McGraw Hill Companies, 2001
Thirteen
Genomics, Biotechnology, and Recombinant DNA
37. Draw the expected gel pattern derived from the dideoxy sequencing method for a template strand with the following sequence: 5 -CAGCGAATGCGGAA-3 38. A DNA strand with the sequence 3 -GACTATTCCGAAAC-5 is sequenced by the dideoxy method. If the reaction mixture contains all four radioactive deoxynucleotide triphosphates plus dideoxythymidine, what size labeled bands do you expect to see on the gel
MAPPING AND SEQUENCING THE HUMAN GENOME
39. What is hypervariable DNA a RFLP a VNTR locus microsatellite DNA a sequence-tagged site (See also RESTRICTION MAPPING)
PRACTICAL BENEFITS FROM GENE CLONING
40. Describe some areas of practical bene t from genetic engineering. Why might some people be concerned about its widespread use
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. In the past, geneticists have used several different methods to splice pieces of DNA that do not have compatible sticky ends. We mentioned blunt-end ligation and the addition of linkers containing speci c restriction sites. Given that nucleotides can be added to the 3 ends of double-stranded DNA with the enzyme deoxynucleoside terminal transferase, can you see an-
other way to create compatible ends on foreign and vehicle DNA 2. The motion picture Jurassic Park was based on the premise that DNA of dinosaurs could be extracted from the blood-meals of mosquitoes preserved in amber and inserted into the genome of a frog, which would then produce living dinosaurs. Is this premise reasonable
Suggested Readings for chapter 13 are on page B-11.
Tamarin: Principles of Genetics, Seventh Edition
III. Molecular Genetics
14. Gene Expression: Control in Prokaryotes and Phages
The McGraw Hill Companies, 2001
GENE EXPRESSION
Control in Prokaryotes and Phages
STUDY OBJECTIVES
1. To study the way in which inducible and repressible operons work 406 2. To examine attenuator control in bacteria 415 418 3. To analyze the control of the life cycle of phage 4. To determine the way in which transposable genetic elements transpose and control gene expression in bacteria 425 5. To look at other transcriptional and posttranscriptional mechanisms of control of gene expression in bacteria and phages 430
STUDY OUTLINE
The Operon Model 406 Lac Operon (Inducible System) 406 Lactose Metabolism 406 The Regulator Gene 406 The Operator 408 Induction of the Lac Operon 409 Lac Operon Mutants 409 Catabolite Repression 412 Trp Operon (Repressible System) 413 Tryptophan Synthesis 413 Operator Control 414 Trp Operon (Attenuator-Controlled System) 415 Leader Transcript 415 Leader Peptide Gene 416 TRAP Control 417 Redundant Controls 418 Lytic and Lysogenic Cycles in Phage 418 Phage Operons 418 Early and Late Transcription 420 Repressor Transcription 421 Maintenance of Repression 421 Lysogenic Versus Lytic Response 423 Transposable Genetic Elements 425 IS Elements 425 Composite Transposons 427 Mechanism of Transposition 427 Phenotypic and Genotypic Effects of Transposition 429 Other Transcriptional Control Systems 430 Transcription Factors 430 Promotor Ef ciency 430 Translational Control 430 Posttranslational Control 433 Feedback Inhibition 433 Protein Degradation 433 Summary 434 Solved Problems 435 Exercises and Problems 436 Critical Thinking Questions 438
Arti cially colored transmission electron micrograph of a T4 bacteriophage attached to an Escherichia coli bacterium. ( Biozentrum, University of Basel/SPL/Photo
Researchers, Inc.)
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