BIOTECHNOLOGY PRICIPLE AND PROCESS

Biotechnology deals with techniques of
using live organisms or enzymes from
organisms to supply products and
processes useful to humans.
• Traditional form – supported natural
capabilities of microorganisms. making
curd, bread or wine, which are all
microbe-mediated processes, could also
be thought as a type of biotechnology.
However, it's employed in a restricted sense
today,
• Modern form – it uses genetically
modified organisms to realize the identical
on a bigger scale. Further, many other

processes/techniques are included
under biotechnology. as an example, in
vitro fertilisation resulting in a ‘test-tube’
baby, synthesising a gene and using it,
developing a DNA vaccine or correcting
a defective gene, are all a part of
biotechnology.
• the eu Federation of
Biotechnology (EFB) has given a
definition of biotechnology that
encompasses both traditional view and
modern molecular biotechnology. The
definition given by EFB is as follows:
‘The integration of scientific discipline
and organisms, cells, parts thereof,
and molecular analogues for
products and services’.
PRINCIPLES OF BIOTECHNOLOGY
• Among many, the 2 core techniques
that enabled birth of recent
biotechnology are :
• Genetic engineering: Techniques to
alter the chemistry of genetic material
(DNA and RNA),to introduce these into
host organisms and thus change the
phenotype of the host organism.
Maintenance of sterile (microbial
contamination-free) ambience in
chemical engineering processes to enable
growth of only the specified
microbe/eukaryotic cell in large quantities
for the manufacture of biotechnological
products like antibiotics, vaccines,
enzymes, etc.
• amphimixis has many
advantages over reproduction.
The former provides opportunities for
variations and formulation of unique
combinations of genetic setup, some of
which may be beneficial to the organism
as well because the population. Asexual
reproduction preserves the genetic
information, while amphimixis
permits variation.
• Traditional hybridisation procedures
used in plant and animal breeding, very
often cause inclusion and
multiplication of undesirable genes
along with the specified genes. Thetechniques of gene-splicing which
include creation of desoxyribonucleic acid,
use of gene cloning and gene transfer,
overcome this limitation and permit us to
isolate and introduce just one or a collection of
desirable genes without introducing
undesirable genes into the target
organism.
• a bit of DNA, which is somehow
transferred into an alien organism, most
likely wouldn't be able to multiply itself
in the progeny cells of the organism.
But, when it gets integrated into the
genome of the recipient, it should multiply
and be inherited together with the host
DNA. this can be because the alien piece of
DNA has become a part of a chromosome,
which has the flexibility to duplicate.
• in an exceedingly chromosome there's a particular DNA
sequence called the origin of replication,
which is to blame for initiating
replication. Therefore, for the
multiplication of any alien piece of DNA
in an organism it has to be a component of a
chromosome(s) which contains a specific
sequence referred to as ‘origin of
replication’. Thus, an alien DNA is
linked with the origin of replication, so
that, this alien piece of DNA can
replicate and multiply itself within the host
organism. this may even be called as
cloning or making multiple identical
copies of any template DNA.
• the development of the primary
recombinant DNA emerged from the
possibility of linking a gene encoding
antibiotic resistance with a native
plasmid (autonomously replicating
circular extra-chromosomal DNA)
of salmonella.
• Stanley Cohen and Herbert Boyer
accomplished this in 1972 by isolating
the antibiotic resistance gene by cutting
out a bit of DNA from a plasmid
which was to blame for conferring
antibiotic The cutting of DNA at specific locations
became possible with the invention of
the so-called ‘molecular scissors’-
restriction enzymes.
• The cut piece of DNA was then linked
with the plasmid DNA. These plasmid
DNA act as vectors to transfer the piece
of DNA attached to that. A plasmid will be
used as vector to deliver an alien piece of
DNA into the host organism.
• The linking of antibiotic resistance gene
with the plasmid vector became possible
with the enzyme DNA ligase, which acts
on cut DNA molecules and joins their
ends. This makes a replacement combination of
circular autonomously replicating DNA
created in vitro and is understood as
recombinant DNA.
• When this DNA is transferred into
Escherichia coli, a bacterium closely
related to Salmonella, it could replicate
using the new host’s DNA polymerase
enzyme and make multiple copies. The.
ability to multiply copies of antibiotic
resistance gene in coli was called cloning
of antibiotic resistance gene in E. coli.
• there are three basic steps in genetically
modifying an organism
• identification of DNA with desirable
genes;
• introduction of the identified DNA into
the host;
• maintenance of introduced DNA within the
host and transfer of the DNA to its
progeny.
TOOLS OF desoxyribonucleic acid
TECHNOLOGY
Key tools of desoxyribonucleic acid technology
are – restriction enzymes, polymerase
enzymes, ligases, vectors and therefore the host
organism.
1. RESTRICTION ENZYMES• In 1963, the 2 enzymes responsible
for restricting the expansion of
bacteriophage in E. coli were
isolated. one in every of these added methyl
groups to DNA, while the opposite cut
DNA. The later was called restriction
endonuclease.
• the primary restriction nuclease
isolated – Hind II.
• restriction nuclease cut DNA
molecules at a selected point by
recognising a particular sequence of base
pairs. This specific base sequence is
known as the popularity
sequence.(For Hind II – sequence of 6
base pairs).
• Today we all know quite 900
restriction enzymes that are
isolated from over 230 strains of
bacteria each of which recognise
different recognition sequences.
Naming of enzymes –• First letter of the name comes from the
genes
• The second two letters come from the
species of the prokaryotic cell from
which they were isolated, e.g., EcoRI