Recombinant DNA technology is the joining together of DNA molecules from two different species. The recombined DNA molecule is inserted into a host organism to produce new genetic combinations that are of value to science, medicine, agriculture, and industry. Since the focus of all genetics is the gene, the fundamental goal of laboratory geneticists is to isolate, characterize, and manipulate genes. Recombinant DNA technology is based primarily on two other technologies, cloning and DNA sequencing. Cloning is undertaken in order to obtain the clone of one particular gene or DNA sequence of interest. The next step after cloning is to find and isolate that clone among other members of the library (a large collection of clones). Once a segment of DNA has been cloned, its nucleotide sequence can be determined. Knowledge of the sequence of a DNA segment has many uses.

Tools of Recombinant DNA Technology

The enzymes which include the restriction enzymes help to cut, the polymerases- help to synthesize and the ligases- help to bind. The restriction enzymes used in recombinant DNA technology play a major role in determining the location at which the desired gene is inserted into the vector genome. They are two types, namely Endonucleases and Exonucleases.

The Endonucleases cut within the DNA strand whereas the Exonucleases remove the nucleotides from the ends of the strands. The restriction endonucleases are sequence-specific which are usually palindrome sequences and cut the DNA at specific points. They scrutinize the length of DNA and make the cut at the specific site called the restriction site. This gives rise to sticky ends in the sequence. The desired genes and the vectors are cut by the same restriction enzymes to obtain the complementary sticky notes, thus making the work of the ligases easy to bind the desired gene to the vector.

The vectors-help in carrying and integrating the desired gene. These form a very important part of the tools of recombinant DNA technology as they are the ultimate vehicles that carry forward the desired gene into the host organism. Plasmids and bacteriophages are the most common vectors in recombinant DNA technology that are used as they have a very high copy number. The vectors are made up of an origin of replication. This is a sequence of nucleotide from where the replication starts, a selectable marker constitute genes which show resistance to certain antibiotics like ampicillin; and cloning sites the sites recognized by the restriction enzymes where desired DNAs are inserted.

Host organism into which the recombinant DNA is introduced. The host is the ultimate tool of recombinant DNA technology which takes in the vector engineered with the desired DNA with the help of the enzymes.

There are a number of ways in which these recombinant DNAs are inserted into the host, namely microinjection, biolistics or gene gun, alternate cooling and heating, use of calcium ions, etc.

Applications of recombinant DNA technology

•          DNA technology is also used to detect the presence of HIV in a person.

•          Gene Therapy-It is used as an attempt to correct the gene defects which give rise to heredity diseases.

•          Clinical diagnosis -ELISA is an example where the application of recombinant

•          Recombinant DNA technology is widely used in Agriculture to produce genetically-modified organisms such as  Flavr Savr tomatoes, golden rice rich in proteins, Bt-cotton to protect the plant against ball worms and lot more.

•          In the field of medicines, Recombinant DNA technology is used for the production of Insulin.

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Nicola B
Editorial Team
Journal of  Biochemistry and Biotechnology