Explain the process of recombinant DNA technology thoroughly with appropriate vocab ?

Recombinant DNA technology is the selection of the desired gene for its administration into the host which is then followed by a selection of the perfect vector with which the gene has to be integrated and recombinant DNA is formed.

Process of recombinant DNA technology.

Isolation of DNA

Isolation of DNA is the first process where it is controlled by enzymes by treating plant or animal cell with an enzyme.Example of enzymes include cellulase for plant cells,chitinase for fungi and lysozyme for bacteria.This enzymes are used in isolating pure DNA from the cells.

Fragmentation of DNA

This is the second process in which the isolated and purified DNA is treated with restriction endonucleases which cuts the DNA into fragments. The restriction enzymes used in recombinant DNA technology helps in determining the location at which the desired gene is inserted into the vector genome,they are also sequence-specific which is palindrome sequences and that it cuts the DNA at specific points. They inspect closely the length of DNA and make the cut at the specific site referred to as the restriction site. The desired genes and the vectors are cut by the same restriction enzymes in order to obtain the complementary sticky ends, hence making the work of the ligases easy to bind the desired gene to the vector.

Amplification of Gene of Interest

Polymerase chain reaction (PCR) is a method that is widely used to making million of copies of a specific DNA sample. Through the polymerase chain reaction multiple copies of the gene of interest can be produced. PCR proceeds in three stages, denaturation, annealing and extension.

Insertion of recombinant DNA into the host.

The host is the ultimate tool of rDNA technology and it takes in the vector engineered with the desired DNA with the help of the enzymes. Insertion of the desired recombinant DNA into the host organism can be achieved through various ways. This includes- gene gun,microinjection, alternate cooling and heating, use of calcium ions, etc. The successfully transformed cells or the organisms carry forward the recombinant gene to the offsprings.