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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
Chapter 14, Problem 13b

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?

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1
Identify a reporter gene that can be used to visually indicate changes in gene expression. A common choice is the GFP (Green Fluorescent Protein) gene, which produces a fluorescent signal that is easily observable under specific light conditions.
Design a genetic construct where the promoter region of the CBF genes (which is activated by low temperatures) is fused to the reporter gene (e.g., GFP). This ensures that the reporter gene is expressed only when the CBF genes are activated by cold conditions.
Introduce the genetic construct into Arabidopsis plants using a method such as Agrobacterium-mediated transformation. This will integrate the construct into the plant genome, allowing the modified plants to express the reporter gene under the control of the CBF promoter.
Grow the genetically modified plants in the field and expose them to low temperatures. Observe the plants under appropriate light conditions (e.g., UV light for GFP) to detect the fluorescence signal, which indicates activation of the CBF genes.
Communicate to farmers that the fluorescence signal in the plants serves as a visual indicator of low-temperature stress, allowing them to monitor environmental conditions and take appropriate agricultural actions if needed.

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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 plants, environmental factors like temperature can influence the expression of specific genes, such as the CBF genes in Arabidopsis, which are involved in cold response. Understanding how gene expression changes in response to low temperatures is crucial for developing methods to observe these changes in a field setting.
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Phenotypic Changes

Phenotypic changes are observable traits or characteristics of an organism that result from the interaction of its genotype with the environment. In the context of Arabidopsis and CBF gene expression, low temperatures may lead to visible changes in plant morphology, such as leaf color or growth patterns. Identifying these phenotypic changes can help farmers recognize the effects of cold stress on their crops.
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Field-Based Observation Techniques

Field-based observation techniques involve methods that allow farmers to assess plant health and stress responses directly in their growing environment. Techniques such as visual assessments of leaf color, growth rate, or the use of simple indicators (like color-coded tags) can help farmers recognize changes in gene expression related to cold exposure. Designing a method that incorporates these techniques can facilitate practical monitoring of plant responses to temperature variations.
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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?

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