Question

In the process of rDNA technology, if two separate restriction enzymes are used to cut vector and donor DNA then which problem will arise in the formation of rDNA or chimeric DNA? Explain.

Answer 1

1. Restriction enzymes cut the DNA at a specific recognition site and produce either sticky ends or blunt ends.
2. For example, if we use the restriction enzyme EcoRI to cut vector DNA, then it will produce vector DNA having sticky ends, and if we use the Hind II restriction enzyme to cut donor DNA, then it will produce donor DNA with blunt ends.
3. A vector DNA to get ligated to the desired gene, they both must possess similar kinds of ends, i.e., both should have either blunt ends or sticky ends.
4. If two separate restriction enzymes are used to cut vector and donor DNA, then they will fail to form complementary base pairing, and chimeric DNA will not be formed.