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?
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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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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