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Ch. 14 - Analysis of Gene Function via Forward Genetics and Reverse Genetics
Sanders - Genetic Analysis: An Integrated Approach 3rd Edition
Sanders3rd EditionGenetic Analysis: An Integrated ApproachISBN: 9780135564172Not the one you use?Change textbook
All textbooksSanders 3rd EditionCh. 14 - Analysis of Gene Function via Forward Genetics and Reverse GeneticsProblem 13a
Chapter 14, Problem 13a

The CBF genes of Arabidopsis are induced by exposure of the plants to low temperature. How would you examine the temporal and spatial patterns of expression after induction by low temperature?

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Design an experiment to monitor the expression of CBF genes over time and in different tissues. This can be achieved by creating transgenic Arabidopsis plants with a reporter gene (e.g., GFP or GUS) fused to the promoter of the CBF genes.
Expose the transgenic plants to low-temperature conditions to induce the expression of the CBF genes. Ensure that the temperature and duration of exposure are consistent across all samples.
Collect tissue samples from different parts of the plant (e.g., leaves, stems, roots) at various time points after the low-temperature exposure. This will allow you to study both temporal (time-based) and spatial (location-based) expression patterns.
Analyze the reporter gene activity in the collected samples. For example, if using GFP, observe fluorescence under a fluorescence microscope. If using GUS, perform a histochemical staining assay to visualize expression patterns.
Quantify the expression levels of the CBF genes using molecular techniques such as quantitative PCR (qPCR) or RNA sequencing (RNA-seq) to validate and complement the reporter gene data. This will provide precise temporal and spatial expression profiles.

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Key Concepts

Here are the essential concepts you must grasp in order to answer the question correctly.

Gene Expression

Gene expression refers to the process by which information from a gene is used to synthesize functional gene products, typically proteins. In the context of the CBF genes in Arabidopsis, understanding how these genes are activated in response to low temperatures is crucial for studying plant responses to environmental stress.
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Temporal and Spatial Patterns

Temporal patterns of gene expression involve measuring how gene activity changes over time, while spatial patterns refer to where in the plant the gene is expressed. Analyzing these patterns helps researchers understand the dynamics of CBF gene activation and its role in cold acclimation in Arabidopsis.
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Induction Mechanisms

Induction mechanisms describe the processes that trigger the activation of specific genes in response to environmental stimuli, such as low temperature. Investigating these mechanisms can reveal the signaling pathways involved in the cold response and how plants adapt to changing climates.
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Related Practice
Textbook Question

Go to the website http://www.ncbi.nlm.nih.gov/omim and enter 'Lynch syndrome' in the Search box at the top of the page. From the list of options given, select '#120435—Lynch Syndrome.' Use the information you retrieve to answer the following questions. Provide a brief summary of the normal functions of the protein products of these genes.

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Textbook Question

You have identified a recessive mutation that alters bristle patterning in Drosophila and have used recombinant DNA technology to identify a genomic clone that you believe harbors the gene. How would you demonstrate that your gene is on the genomic clone?

Textbook Question

Genetic counseling has not been discussed in this chapter, but it is a service provided by trained professional counselors who also have detailed knowledge of medical genetics, as described in Application Chapter A. Genetic counselors provide details about gene mutations and have knowledge of most of the details of diseases associated with genetic abnormalities. With regard to genetic testing to identify one's personal risk of cancer, what are the three or four topics you think are most important to be able to discuss with a genetic counselor?

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Textbook Question

The CBF genes of Arabidopsis are induced by exposure of the plants to low temperature. Can you design a method that would reveal these changes in gene expression in a way that a farmer could recognize them by observing plants growing in the field?

Textbook Question

When the S. cerevisiae genome was sequenced and surveyed for possible genes, only about 40% of those genes had been previously identified in forward genetic screens. This left about 60% of predicted genes with no known function, leading some to dub the genes fun (function unknown) genes. As an approach to understanding the function of a certain fun gene, you wish to create a loss-of-function allele. How will you accomplish this?

Textbook Question

When the S. cerevisiae genome was sequenced and surveyed for possible genes, only about 40% of those genes had been previously identified in forward genetic screens. This left about 60% of predicted genes with no known function, leading some to dub the genes fun (function unknown) genes. You wish to know the physical location of the encoded protein product. How will you obtain such information?