Question
i. Explain how to find whether an E.coli bacterium has transformed or not when a recombinant DNA bearing ampicillin resistant gene is transferred into it.
ii. What does the ampicillin resistant gene act as in the above case?

Answer

i. When an E.coli bacterium is transformed with a recombinant DNA bearing ampicillin-resistant gene in its plasmid, the recombinant plasmid will lose tetracycline resistance due to the insertion of foreign DNA but can still be selected out from non-recombinant ones by plating the transformants on ampicillin containing a medium. The transformants growing on ampicillin containing medium are then transferred to a medium containing tetracycline. The recombinants will grow on ampicillin containing medium but not one that containing tetracycline. But non-recombinant will grow on the medium containing both the antibiotics.
ii. In this case, one antibiotic gene helps in selecting the transformants whereas the other antibiotic resistance gene gets inactivated due to the insertion of alien DNA and helps in the selection of recombinants. Ampicillin resistant gene in the above case helps in selecting the transformants and act as a selectable marker.

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