Plant Biotechnology -16 || Tools of rDNA Technology || Gene of interest || Vector || Enzyme || Host

Tools of rDNA Technology
There are four major requirements of gene cloning or recombinant DNA Technology.
Gene of interest
Vector molecule
Enzyme
Host
cloning vector in biotechnology
what is gene of interest?
The DNA sequence that has a desirable phenotypic expression i.e., genes for biotic and stress resistance, genes for quality traits etc.
The DNA sequence that is to be cloned.
The DNA sequence that is to be transferred for production of transgenic or genetically modified organism (Transgene).
The DNA sequence that is to be transferred from one organism to another organism in which it is not occurs naturally (foreign gene).
What are vector?
Vectors are the carriers of genes which carry the genes from one organism to another organism and replicate it into many copies.
Vector is a recombinant DNA molecule that is capable of independent replication when introduced into a suitable organism.
Vector acts as a vehicle that transports the gene into a host cell (usually, bacterium).
Ex. Plasmid, cosmid, phagemid, YAC and BAC etc.
What are restriction endonucleases?
Endonucleases are the enzymes that produce internal cuts, called cleavage (both the strand are cut) in DNA molecule.

The endonuclease enzyme that cut the DNA at or near a specific sequence are termed as restriction endonuclease enzymes.
The site recognized by the enzymes is known as recognition site or recognition sequence.
Restriction enzymes are also known as Molecular Scissors
What are ligases?
DNA ligase is a enzyme that can link together DNA strands that have double-strand breaks (a break in both complementary strands of DNA).
Naturally DNA ligase has applications in both DNA replication and DNA repair .
Needs ATP
DNA ligase has extensive use in molecular biology laboratories for genetic recombination experiments
What is host?
Any organism (usually bacterium) in which a vector molecule can multiply.
Properties of a good host:
It should be easy to transform.
It should support the replication of recombinant DNA.
It should be free from the elements that interfere with replication of recombinant DNA.

It should lack active restriction enzyme e.g., Ecoli, K12, HB 101.
It should not have methylase. (methylase protect DNA from RE).
It should be deficient in normal recombination, so that the DNA insert is not altered.

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