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6. Suppose a particular gene is required for early development and also later for development of a particular tissue, such as the adult nervous system. By generating a homozygous mutant clone in that tissue of a heterozygote, researchers can circumvent the lethality that would result if the entire animal is homozygous for a loss-of-function mutation in that gene. A technique called MARCM (Mosaic Analysis with a Repressible Cell Marker) was developed to enable Drosophila geneticists to generate homozygous mutant cell clones that are marked by the presence of a reporter protein such as GFP. Marker expression enables the investigator to observe clearly the mutant phenotype within a clone of mutant cells. This technique relies on a yeast protein called Gal80 that is a negative regulator of the Gal4 protein described previously in Solved Problem II. Gal80 binds to Gal4 and prevents it from activating transcription. The idea of MARCM is that Gal4/UASG-driven GFP expression is blocked by Gal80 throughout the fly, except within the homozygous mutant clone where the Gal80-expressing transgene is lost by mitotic recombination. a. Diagram the chromosomes and the mitotic crossover that generate a homozygous m- mutant clone marked by GFP expression. b. How could you restrict the clones to the adult nervous system?

6. Suppose a particular gene is required for early development and also later for development of a particular tissue, such as the adult nervous system. By generating a homozygous mutant clone in that tissue of a heterozygote, researchers can circumvent the lethality that would result if the entire animal is homozygous for a loss-of-function mutation in that gene. A technique called MARCM (Mosaic Analysis with a Repressible Cell Marker) was developed to enable Drosophila geneticists to generate homozygous mutant cell clones that are marked by the presence of a reporter protein such as GFP. Marker expression enables the investigator to observe clearly the mutant phenotype within a clone of mutant cells. This technique relies on a yeast protein called Gal80 that is a negative regulator of the Gal4 protein described previously in Solved Problem II. Gal80 binds to Gal4 and prevents it from activating transcription. The idea of MARCM is that Gal4/UASG-driven GFP expression is blocked by Gal80 throughout the fly, except within the homozygous mutant clone where the Gal80-expressing transgene is lost by mitotic recombination. a. Diagram the chromosomes and the mitotic crossover that generate a homozygous m- mutant clone marked by GFP expression. b. How could you restrict the clones to the adult nervous system?

Biology: The Dynamic Science (MindTap Course List)
Biology: The Dynamic Science (MindTap Course List)
4th Edition
ISBN:9781305389892
Author:Peter J. Russell, Paul E. Hertz, Beverly McMillan
Publisher:Peter J. Russell, Paul E. Hertz, Beverly McMillan
Chapter16: Regulation Of Gene Expression
Section: Chapter Questions
Problem 8TYK
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6. Suppose a particular gene is required for early development and also later for development of a particular tissue, such as the adult nervous system. By generating a homozygous mutant clone in that tissue of a heterozygote, researchers can circumvent the lethality that would result if the entire animal is homozygous for a loss-of-function mutation in that gene. A technique called MARCM (Mosaic Analysis with a Repressible Cell Marker) was developed to enable Drosophila geneticists to generate homozygous mutant cell clones that are marked by the presence of a reporter protein such as GFP. Marker expression enables the investigator to observe clearly the mutant phenotype within a clone of mutant cells. This technique relies on a yeast protein called Gal80 that is a negative regulator of the Gal4 protein described previously in Solved Problem II. Gal80 binds to Gal4 and prevents it from activating transcription. The idea of MARCM is that Gal4/UASG-driven GFP expression is blocked by Gal80 throughout the fly, except within the homozygous mutant clone where the Gal80-expressing transgene is lost by mitotic recombination. a. Diagram the chromosomes and the mitotic crossover that generate a homozygous m- mutant clone marked by GFP expression. b. How could you restrict the clones to the adult nervous system?
Transcribed Image Text:6. Suppose a particular gene is required for early development and also later for development of a particular tissue, such as the adult nervous system. By generating a homozygous mutant clone in that tissue of a heterozygote, researchers can circumvent the lethality that would result if the entire animal is homozygous for a loss-of-function mutation in that gene. A technique called MARCM (Mosaic Analysis with a Repressible Cell Marker) was developed to enable Drosophila geneticists to generate homozygous mutant cell clones that are marked by the presence of a reporter protein such as GFP. Marker expression enables the investigator to observe clearly the mutant phenotype within a clone of mutant cells. This technique relies on a yeast protein called Gal80 that is a negative regulator of the Gal4 protein described previously in Solved Problem II. Gal80 binds to Gal4 and prevents it from activating transcription. The idea of MARCM is that Gal4/UASG-driven GFP expression is blocked by Gal80 throughout the fly, except within the homozygous mutant clone where the Gal80-expressing transgene is lost by mitotic recombination. a. Diagram the chromosomes and the mitotic crossover that generate a homozygous m- mutant clone marked by GFP expression. b. How could you restrict the clones to the adult nervous system?
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