Based on what you now know about bacterial transformation pr

Based on what you now know about bacterial transformation, predict the impact of inserting a very large foreign DNA fragment (5+ Kb) into pUC18 on….

a. the number of plasmids replicated in each cell

b. the efficiency of transformation (i.e., the percentage of cells that took up the plasmid)

c. the phenotype (color) of the colonies transformed with the plasmid.

Solution

a. the number of plasmids replicated in each cell

The puc18 is a vector that is of 2686 bp. So l inserting copies up to 10kb can be possible in a pUC18 vector as it is a cloning vector. The DNA will be cloned after insert within this plasmid. One copy of the DNA will be formed unless cloned further.

b. the efficiency of transformation (i.e., the percentage of cells that took up the plasmid)

Cannot be guessed, the transformation efficiency depends on the different cells.

c. the phenotype (color) of the colonies transformed with the plasmid.

One of the primary features of the pUC18 vector is its ability of distinguishing the recombinant and non recombinant Cells via colour production. Due to the presence of beta galactosidase in its multiple cloning site, on reaction with X- Gal and IPTG, THERE IS A BLUE COLOUR, production, which will be white in case of recombinant colonies. Only the cells which have taken up the plasmid containing the amp^R gene will survive.