You have amplified the cDNA sequence for the ‘Protein A’ gene using PCR amplification. Now you wish to splice the gene into a plasmid in order to transform E.
coli with the plasmid. Your plasmid contains an ampR gene. The plasmid also has a bgalactosidase (lacZ) gene. The lacZ gene contains an EcoRI restriction enzyme site.
After you digest the PCR products and your plasmids with EcoRI restriction enzyme,
explain
a. how you would insert the ‘Protein A’ gene into the plasmid;
b. after transforming cells with the plasmids, how you would screen for colonies of cells
that had incorporated a plasmid;
c. after transforming the cells with plasmids, how you would screen for colonies of cells
that had taken on recombinant plasmids (i. e. plasmids with the ‘Protein A’ PCR product);
d. and finally, explain how and why the colonies that have taken on the insert are
different from colonies that were transformed with plasmids that did not take on the
‘Protein A’ gene insert