[figure] In the above represented plasmid, an alien piece of DNA is inserted at EcoRI site. Which of the following strategies will be chosen to select the recombinant colonies?
- White color colonies will be selected.
- Blue color colonies grown on ampicillin plates can be selected.
- Using ampicillin & tetracycline containing medium plate.
- Blue color colonies will be selected.
Answer
(1) White color colonies will be selected.
Solution
The EcoRI site lies inside the coding sequence of the enzyme -galactosidase.
Inserting alien DNA at this site causes insertional inactivation of the -galactosidase gene.
On a medium containing a chromogenic substrate, non-recombinant colonies (active -galactosidase) give blue colour, while recombinant colonies (inactivated gene) give no colour, i.e. they appear white.
Hence, the white colonies are selected as recombinants.
Answer: (1)