Formation of Recombinant DNA
- Recombinant DNA is formed by using a restriction enzyme that cuts the double strand at a particular point.
- The same enzyme is used to cut a second piece of DNA.
- When the fragments are mixed together, the complementary ends of each strand will bind with those of the other, forming a recombinant DNA molecule.
Process of formation of Recombinant DNA
The process of recombination DNA technology consists of the following steps:
- Isolation of genetic material (DNA)
- DNA is enclosed within the membrane. So, to extract it out, the cell wall is broken down to release the genetic material.
- This can be done by using cell wall digesting enzymes such as Lysozymes for bacterial cells, cellulose for plant cells and chitinase for fungi cells.
- Cutting of DNA at specific locations
- This is done by using restriction enzymes that cut DNA at specific recognition sequences and give the desired DNA fragments.
- Joining of DNA fragment
- The DNA fragments generated with a cloning vector can be done by sticky ends ligation, blunt-end ligation and homopolymer tailing.
- Insertion of DNA into the host cell.
- The recombinant DNA can be inserted into the host cell by transformation, transduction, and vectorless gene transfer.
- Selection and screening of transformed cells
- The selection and screening of transformed cells can be by any of the following methods- immunological method, nucleic acid hybridization and blue-white screening or insertional inactivation.