During the analysis of the genomic sequence of the mouse genome, you have found a gene...

During the analysis of the genomic sequence of the mouse genome, you have found a gene that you suspect may be involved in resistance of infection to bubonic plague. You want to test your suspicions by creating a knockout mouse. Outline the steps you would have go through to create such a mouse that is homozygouse for disruption of both copies of this gene. Plese include terms like ES cells, neomycin, positive-negative selction, homologous recombination, blastocyst, and chimera.

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to knocked out gene involved bubonic plague the gene will be dirsrupted or replaced so that the gene is no more functional.

At the begiing the gene for bubonic plague.will be isolated from a gene library which is specific for mouse. Next this DNA sequence will be genetically modified so that this sequence can not any more produce the same protein which is required for the bubonic plague. infection. Additionally a marker gene for eg. an antibiotic resistance cassette such as neomycin resistant gene also introduced in this sequence so that the mice which will have the recombinant construct can be selected.

Next, embryonic stem cells are isolated from a young mouse, which is called blastocyst and will be grown in invitro condition.

After this step, the recombinant DNA will be incorporated into the embyronic stem cells by specialized methods like electroporation. Through the process of hoomlogous recombination, some of the embryonic stem cell will take up the recombinant DNA and their original sequence required for bubonic plague. expression will be relaced by the recombinant cassette.

The embryonic stem cells which has the recombinant cassette will be further chosen on the basis of neomycin selecion marker. The ES cells which have the recombinant cassette will only be able to grow in presence of neomycin.

After this ES cells will be introduced into blastocyst and the blastocysts will be placed into the uterus of a mature mouse and the newborn generated from this female mouse will be called as chimera as they contain both the stem cells from original and also the recombinant one.

When this chimera mouse are crossbreed with wild type mouse some of the newborns will contain of the knocked out gene of bubonic plague.

gladiator answered 1 year ago

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