Textbook Question
Compare and contrast methods for making transgenic plants and transgenic Drosophila.
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Ch. 15 - Recombinant DNA Technology and Its Applications
Sanders3rd EditionGenetic Analysis: An Integrated ApproachISBN: 9780135564172
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Table of contents
- Ch. 1 - The Molecular Basis of Heredity, Variation, and Evolution64
- Ch. 2 - Transmission Genetics144
- Ch. 3 - Cell Division and Chromosome Heredity81
- Ch. 4 - Gene Interaction87
- Ch. 5 - Genetic Linkage and Mapping in Eukaryotes103
- Ch. 6 - Genetic Analysis and Mapping in Bacteria and Bacteriophages73
- Ch. 7 - DNA Structure and Replication71
- Ch. 8 - Molecular Biology of Transcription and RNA Processing79
- Ch. 9 - The Molecular Biology of Translation110
- Ch. 10 - Eukaryotic Chromosome Abnormalities and Molecular Organization100
- Ch. 11 - Gene Mutation, DNA Repair, and Homologous Recombination86
- Ch. 12 - Regulation of Gene Expression in Bacteria and Bacteriophage90
- Ch. 13 - Regulation of Gene Expression in Eukaryotes38
- Ch. 14 - Analysis of Gene Function via Forward Genetics and Reverse Genetics72
- Ch. 15 - Recombinant DNA Technology and Its Applications69
- Ch. 16 - Genomics: Genetics from a Whole-Genome Perspective49
- Ch. 17 - Organelle Inheritance and the Evolution of Organelle Genomes27
- Ch. 18 - Developmental Genetics43
- Ch. 19 - Genetic Analysis of Quantitative Traits66
- Ch. 20 - Population Genetics and Evolution at the Population, Species, and Molecular Levels105
Sanders3rd EditionGenetic Analysis: An Integrated ApproachISBN: 9780135564172Not the one you use?Change textbook
Chapter 15, Problem 15a
The bacteriophage lambda genome can exist in either a linear form or a circular form.
How many fragments will be formed by restriction enzyme digestion with XhoI alone, with XbaI alone, and with both XhoI and XbaI in the linear and circular forms of the lambda genome?
Verified step by step guidance1
Step 1: Understand the lambda genome structure. The bacteriophage lambda genome can exist in two forms: linear and circular. In the linear form, the genome has free ends, while in the circular form, the ends are joined to form a loop. This structural difference affects how restriction enzymes cut the DNA.
Step 2: Learn about the restriction enzymes XhoI and XbaI. Restriction enzymes recognize specific DNA sequences and cut at those sites. For example, XhoI recognizes the sequence 5'-CTCGAG-3', and XbaI recognizes the sequence 5'-TCTAGA-3'. The number of fragments produced depends on the number of recognition sites in the genome.
Step 3: Determine the number of recognition sites for XhoI and XbaI in the lambda genome. This information is typically provided in the genome map or sequence data. Count the number of sites for each enzyme in both the linear and circular forms of the genome.
Step 4: Analyze the cutting pattern for each enzyme. In the linear form, each recognition site results in a cut that produces two fragments. In the circular form, the genome is a loop, so the number of fragments produced is equal to the number of cuts made by the enzyme.
Step 5: Combine the effects of both enzymes. When both XhoI and XbaI are used together, the total number of fragments is determined by the combined number of cuts made by both enzymes. Consider how overlapping recognition sites (if any) might affect the fragment count in both linear and circular forms.
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Key Concepts
Here are the essential concepts you must grasp in order to answer the question correctly.
Bacteriophage Lambda Genome Structure
The bacteriophage lambda genome can exist in two forms: linear and circular. The linear form is typically found during the infection process, while the circular form can occur after the genome is integrated into the host's DNA. Understanding these forms is crucial for predicting how the genome will behave during restriction enzyme digestion.
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Bacteriophage Life Cycle
Restriction Enzymes
Restriction enzymes, such as XhoI and XbaI, are proteins that cut DNA at specific sequences. XhoI recognizes the sequence CTCGAG, while XbaI recognizes the sequence TCTAGA. The number of fragments produced by digestion depends on the number of recognition sites present in the DNA, which varies between the linear and circular forms of the genome.
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DNA Fragmentation and Analysis
When a DNA molecule is cut by restriction enzymes, it is fragmented into smaller pieces. The number of fragments generated can be calculated based on the number of cutting sites. In the case of the lambda genome, analyzing the resulting fragments from different enzyme combinations helps in understanding the genome's structure and the effects of the enzymes used.
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Related Practice
Textbook Question
It is often desirable to insert cDNAs into a cloning vector in such a way that all the cDNA clones will have the same orientation with respect to the sequences of the plasmid. This is referred to as directional cloning. Outline how you would directionally clone a cDNA library in the plasmid vector pUC18.
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Textbook Question
A major advance in the 1980s was the development of technology to synthesize short oligonucleotides. This work both facilitated DNA sequencing and led to the advent of the development of PCR. Recently, rapid advances have occurred in the technology to chemically synthesize DNA, and sequences up to 10 kb are now readily produced. As this process becomes more economical, how will it affect the gene-cloning approaches outlined in this chapter? In other words, what types of techniques does this new technology have potential to supplant, and what techniques will not be affected by it?
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Textbook Question
The bacteriophage lambda genome can exist in either a linear form or a circular form.
Diagram the resulting fragments as they would appear on an agarose gel after electrophoresis.
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Textbook Question
The restriction enzymes XhoI and SalI cut their specific sequences as shown below:
Can the sticky ends created by XhoI and SalI sites be ligated? If yes, can the resulting sequences be cleaved by either XhoI or SalI?
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Textbook Question
The bacteriophage ϕX174 has a single-stranded DNA genome of 5386 bases. During DNA replication, double-stranded forms of the genome are generated. In an effort to create a restriction map of ϕX174, you digest the z-stranded form of the genome with several restriction enzymes and obtain the following results. Draw a map of the ϕX174 genome.
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