This document discusses biotechnology and recombinant DNA technology. It defines biotechnology as using organisms or enzymes to produce useful products. It explains the principles of genetic engineering and maintaining sterile environments. It also describes the basic tools used in recombinant DNA technology, including restriction endonucleases, cloning vectors, competent host cells. Finally, it outlines the processes involved in recombinant DNA technology such as isolating genetic material, inserting DNA into host cells, and using bioreactors to produce products at large scale.

2. Definition of Biotechnology
Principles of Biotechnology
Tools of Recombinant DNA technology
Processes of Recombinant DNA technology

3. Definition of Biotechnology
Biotechnology deals with techniques
of using live organisms or enzymes
from organisms to produce products
and processes useful to humans

4. Definition by
European Federation of Biotechnology (EFB)
The integration of Natural science
and organisms, cells, parts thereof
and molecular anlogues
for products and services

5. Principles of Biotechnology
Genetic
Engineering
Maintenance of
sterile
Environment

6. 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

7. Maintenance of sterile environment
Maintenance of sterile ambience in chemical engineering
processes to enable growth of only the desired microbe/
eukaryotic cell in large quantities for the manufacture of
biotechnological products like antibiotics, vaccines, enzymes etc.

8. 3 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 in the host and transfer of the
DNA to its progeny

9. Tools of Biotechnology
Restriction
Endonuclease
Enzyme
Cloning Vectors Competent Host

10. 1963: 2 Enzymes are responsible for restricting the growth of bacteriophage in
Escherichia coli were isolated
The enzyme which adds methyl groups: Deoxyadenosine methylase (DAM)
The enzyme which cuts DNA: Restriction Endonuclease
First endonuclease found was Hind II
Eco RI
Eco: Escherichia coli (Name of the bacteria)
R: RY strain
I: Roman number indicating the order in which enzyme was isolated from the
bacteria
RE recognizes palindromic sequences: DNA sequence of base pairs that reads
same on the 2 strands when orientation of reading is kept the same.

14. Features required to facilitate Cloning in
Vector
Origin of
replication (Ori)
Selectable
marker
Cloning Sites
Vectors for
cloning in Plants
and Animals
Vectors used for preparing recombinant DNA (Inserting DNA of
our choice/ desired gene) are known as Cloning Vectors

15. Sequence from where replication starts is known as Ori
This sequence is also responsible for
controlling the copy number of the linked
DNA.
So if many copies are to be recovered the
vector must have high copy number
Any piece of DNA when linked to this sequence can be made to replicate
within the host cells.

17. Site where DNA to be cloned is inserted
in the vector
This is usually a restriction enzyme site
present in any of the selectable marker
site
Ex: BamH I restriction enzyme site is the
cloning site for the selectable marker
tetracyclin

18. Selection of the recombinant is done by insertional inactivation:
Inactivation of the removed gene due to new insert

21. In order to force bacteria to take up the plasmid the bacterial cells must first be
made competent by treating them with calcium ions

22. Methods of introduction of rDNA in Host
Heat Shock Microinjection
Bollistic
Method
Use of disarmed
pathogen

26. Process of Recombinant
DNA technology
Isolation
of genetic
material
Cutting of
DNA at
specific
location
Amplificati
on of gene
of interest
using PCR
Insertion of
recombinant
DNA into the
host cell/
organism
Obtaining
the foreign
gene
product

27. Lysozyme Cellulase Chitinase
Bacterial
cells
Plant
cells
Animal
cells
Ribonucleasese
+ Proteases
Isolated DNA

31. Bioreactors are used to yield appreciable quantities of product to
produce in large quantities

35. The processes like separation and purification of biosynthetic
product before it is ready for marketing as a finished product is
known as Downstream processing
Steps of Downstream processing
Formulation of product with preservatives
Clinical trials of drugs/ formulations
Strict quality control testing for each product