chapter_14
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| chapter_14 [2025/04/29 11:55] – [Applying the cis/trans test in eukaryotic gene regulation] mike | chapter_14 [2025/04/29 11:56] (current) – [Introduction] mike | ||
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| In this chapter we will see how genetics can be used to dissect molecular structure and function. We have seen one example of how genetics can achieve this in [[Chapter_13|Chapter 13]] with the $gal4^{81}$mutant - this mutant taught us something about how Gal80p functions in relation to Gal4p. In this chapter we will look at two further examples of how genetics can teach us about molecular function. In our first example, we will look at a genetic approach that can be used to analyze the DNA of upstream regulatory sequences of a gene such as $GAL1$. In our second example, we will look at how we can use genetics to assign biochemical functions to different parts of a protein. We will start to think about a new genetic approach to studying biological function: reverse genetics. | In this chapter we will see how genetics can be used to dissect molecular structure and function. We have seen one example of how genetics can achieve this in [[Chapter_13|Chapter 13]] with the $gal4^{81}$mutant - this mutant taught us something about how Gal80p functions in relation to Gal4p. In this chapter we will look at two further examples of how genetics can teach us about molecular function. In our first example, we will look at a genetic approach that can be used to analyze the DNA of upstream regulatory sequences of a gene such as $GAL1$. In our second example, we will look at how we can use genetics to assign biochemical functions to different parts of a protein. We will start to think about a new genetic approach to studying biological function: reverse genetics. | ||
| - | Reverse genetics is the opposite of what we have been studying so far. In Chapters 1-13, we discussed mutants with interesting phenotypes, and used mapping and cloning by complementation to try to identify the protein coding sequence of the gene that is mutated - this approach starts with function (as defined by mutants) and ends with identifying a gene, and is is called forward genetics. In reverse genetics, we already have a gene that has been cloned. We might know of this gene from a whole genome sequencing project; or we might have found a mouse gene based on sequence similarity to a Drosophila gene that was found by forward genetics. We are interested in studying the function of this newly discovered gene. In some cases, the function of the gene is completely unknown. In other cases, we may already know the normal function of a gene, but we want to modify it in some way to further understand details of how the gene product works. In essence, reverse genetics starts with a gene and ends with identifying a function. | + | Reverse genetics is the opposite of what we have been studying so far. In Chapters 1-13, we discussed mutants with interesting phenotypes, and used mapping and cloning by complementation to try to identify the protein coding sequence of the gene that is mutated - this approach starts with function (as defined by mutants) and ends with identifying a gene, and is called forward genetics. In reverse genetics, we already have a gene that has been cloned. We might know of this gene from a whole genome sequencing project; or we might have found a mouse gene based on sequence similarity to a Drosophila gene that was found by forward genetics. We are interested in studying the function of this newly discovered gene. In some cases, the function of the gene is completely unknown. In other cases, we may already know the normal function of a gene, but we want to modify it in some way to further understand details of how the gene product works. In essence, reverse genetics starts with a gene and ends with identifying a function. |
| In this chapter we will discuss three examples of reverse genetic strategies in yeast to study gene function: (1) using reporter genes to study cis-acting regulatory sequences; (2) using the yeast two hybrid system to study protein-protein interactions; | In this chapter we will discuss three examples of reverse genetic strategies in yeast to study gene function: (1) using reporter genes to study cis-acting regulatory sequences; (2) using the yeast two hybrid system to study protein-protein interactions; | ||
chapter_14.1745952913.txt.gz · Last modified: 2025/04/29 11:55 by mike