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Part A

Many cloning vectors have built in antibiotic resistance$$ genes, which are used for selection of the bacteria that picked up the plasmid. The antibiotic resistance$$ gene, allows the bacteria to grow in media containing the antibiotic. Bacteria that fail to pick up the$$ plasmid, cannot grow in the antibiotic containing media.

Suppose that you have an EcoRI recognition site within the antibiotic resistance gene. This restriction$$ site, is the only EcoRI site on the vector. For your research you would like to introduce a fluorescence gene into E$.$

colibacteria.

In order to do$$ this, you digest the fluorescence gene with EcoRI as EcoRI sites are found on either side of your insert and ligate this fragment into the vector$($also cut with$$ EcoRI$).$ Subsequently the bacteria were transformed with this vector. Which of the following statements are$$ TRUE?

I. The fluorescence gene cut with the restriction enzyme EcoRI should be able to get inserted into the plasmid cut with the restriction enzyme

EcoRI.

II$.$ The fluorescence gene cut with the restriction enzyme EcoRI would not be able to get inserted into the plasmid cut with the restriction enzyme

EcoRI.

III. The fluorescence gene will interrupt the normal sequence of the antibiotic resistance gene.

IV$.$ Cells that pick up the plasmid with the fluorescence gene should grow in antibiotic containing media.

V$.$ Cells that pick up the plasmid with the fluorescence gene should not grow in antibiotic containing media.