Question

Enlist the basic steps involved in recombinant DNA technology.

Answer 1

The recombinant DNA molecules formed by the genetic recombination to get the genetic material from the multiple sources. The recombinant DNA is the common name for a piece of DNA that has been produced by combining two fragments from the two different sources.

Complete answer:
Recombinant DNA is the DNA which consists of sequences which are obtained from different sources. The process of recombination of DNA technology includes the following processes:
1. Isolation of DNA : DNA is surrounded by a membrane and to remove it out, the cell wall is broken down to release the genetic material. This process can take place with the help Lysozymes which is the digestive enzymes of the cell membrane for bacterial cells, cellulose for the plant cells and chitinase for the fungi cells.
2. Cutting of DNA at specific locations : This process is done by using restriction enzymes E.coliRI which helps in the cutting sequence of DNA at a particular recognition sequence and gives the desired DNA fragments.
3. Joining of DNA fragments : The DNA fragments can be generated by a cloning vector which can be done by the process sticky ends ligation, blunt-end ligation and homopolymer tailing.
4. Insertion of DNA into the host cell : The desired recombinant DNA can be inserted into the host cell by the process like transformation, transduction, and vectors gene transfer.
5. Selection and screening of transformed cells : The transformed cells are then selected and screened by the following immunological process such as nucleic acid hybridization and blue-white screening or insertional inactivation.

Note: The recombinant DNA is different from genetic recombination in such a way that the recombinant DNA is produced by the artificial method while the genetic recombinant is a biological process. The proteins that are produced by the recombinant DNA within a living cell are known as recombinant proteins.