Question: Suppose you are engineering a mouse with a homozygous knockout
of a gene M as discussed in chapte…
suppose you are engineering a mouse with a homozygous knockout
of a gene M as discussed in chapter 10 (gene replacement). You have
completed many steps in the process and now, you are screening
mouse pups for those that are heterozygous for the knockout – i.e,
mice that have the wild type M allele on one chromosome, and the
disrupted m allele on the other chromosome. To test a given mouse,
you extract its genomic DNA, cut it with EcoRI, and perform a
southern blot. A diagram of the wild type and knockout chromosomes
is shown below. The position where the probe hybridizes is also
shown.
a) your southern analysis includes mice of all possible
genotypes – MM, Mm, and mm. Draw the banding pattern you would
expect for each genotype on the blot A below.
b) Suppose the probe hybridizes only to the inserted DNA ( the
grey box on m chromosome). Draw the banding pattern you would
expect for each genotype on blot B above.
Show transcribed image text 19. (10 points) Suppose you are engineering a mouse with a homozygous knockout of gene M as discussed in chapter 10 (gene replacement). You have completed many steps in the process and now, you are screening mouse pups for those that are heterozygous for the knockout i.e., mice that have the wild type Mallele on one chromosome, and the disrupted m allele on the other chromosome. To test a given mouse, you extract its genomic DNA, cut it with EcoRI, and perform a southern blot. A diagram of the wild type and knockout chromosomes is shown below The position where the probe hybridizes is also shown. 6 kb EcoRI EcoRI 8 kb EcoRl EcoRI Probe: – (a) Your southern analysis includes mice of all possible genotypes-MM, Mm, and mm. Draw the banding pattern you would expect for each genotype on blot A below 8 kb 8 kb 6 kb 6 kb BLOT A BLOTB (b) Suppose the probe hybridizes only to the inserted DNA (the grey box on m chromosome). Draw the banding pattern you would expect for each genotype on blot B above.
19. (10 points) Suppose you are engineering a mouse with a homozygous knockout of gene M as discussed in chapter 10 (gene replacement). You have completed many steps in the process and now, you are screening mouse pups for those that are heterozygous for the knockout i.e., mice that have the wild type Mallele on one chromosome, and the disrupted m allele on the other chromosome. To test a given mouse, you extract its genomic DNA, cut it with EcoRI, and perform a southern blot. A diagram of the wild type and knockout chromosomes is shown below The position where the probe hybridizes is also shown. 6 kb EcoRI EcoRI 8 kb EcoRl EcoRI Probe: – (a) Your southern analysis includes mice of all possible genotypes-MM, Mm, and mm. Draw the banding pattern you would expect for each genotype on blot A below 8 kb 8 kb 6 kb 6 kb BLOT A BLOTB (b) Suppose the probe hybridizes only to the inserted DNA (the grey box on m chromosome). Draw the banding pattern you would expect for each genotype on blot B above.
