6. Principles of Genetic Engineering.pdf

This slide is about genetic engineering

1. Principles of Genetic Engineering

2. Genetic engineering ;-
It is a set of techniques that is used to achieve one or more goals :-
To reveal the processes of how genes are inherited and expressed ……….
To provide effective treatment for various diseases ( mainly genetic disorders)……..
To generate economic benefits which include improved plants and animals for agriculture , and
efficient production of valuable biopharmaceuticals…….

3. Key terms
rDNA
TECHNOLOGY
Gene
therapy
Genetic
engineering
PCR
GMOs
Gene

4. Genetic engineering –
it involves deliberate
DNA manipulation in
organisms to alter
their genes.
Itinvolvesallbiomedicaltechniqueslike-AI,in
vitrofertilization,cloning,genemanipulation
AndrDNATech.etc.
Geneticengineeringbasedonrecombination
waspioneeredin1973byStanleyN.Cohenand
HerbertW.Boyer(Americanbiochemist),who
wasfirstcutDNAintofragmentsandrejoining
andinsertedintoE.coli,whichthenreproduce.

5. Basic principles -

6. 1. Location and
isolation of gene of
interest
2.Insertion of a gene
into a cloning vector
3. Transfer the
recombinant vector
into bacterial cells by
transformation or
infection with virus.
4. Selection of cells
containing the desired
recombinant vectors.
5. Growth of the
bacteria, that can be
continued to give
required cloned DNA
6. Expression of the
gene to get the
desired product.

7. molecular scissors

9. Applications of genetic engineering -
Production of pharmaceutical products
Recombinant vaccines
Diagnosis and cure of diseases
Gene therapy
Transgenic plants and animals
Interferons, hormones ( insulin)

10. Transgenic of genetically modified organisms

12. Diagnosis and cure -

13. Interferons-
Human interferons are inserted into E.coli for production of
interferons by rDNA technology. The complimentary DNA Wes
synthesized from mRNA of specific interferon.
This is inserted into a plasmid vector and then
transferred to the cells (yeast)
The DNA sequence coding for human leukocyte interferon is
attached to the yeast alcohol dehydrogenase gene in
plasmid. Yeast used –saccharomyces cerevisiae.

14. Vaccine production –( hepatitis B vaccine having surface
antigen -: Hbs Ag)
Genetic material is extracted from hepatitis B virus and
individual genes are analysed and identified, genes which
direct produce surface proteins is located and removed from
viral DNA.
This gene is inserted into plasmid and plasmids are inserted
into yeast cells →grown in fermenters →cells are harvested
and disrupts to release the recombinant surface antigen →
purified.
Surface proteins are combined with preserving agents to
make vaccines.
First recombinant vaccine was developed in India by
shantha biotechnics pvt. Ltd. Hyderabad in 1997.

15. Insulin production -
Insulin was first prepared by rDNA technology
and mRNA from rat pancreas by Itakura et al
(1977).
Chemically synthesized A and B chains ⛓️of
insulin are separately inserted into pBR322
plasmid by side of B-galactosidase gene .
The recombinant plasmids are separately
transferred into E.coli cells →grown into
fermenters →two chains are detaches from
B-gactosidase by cynogen bromide →
joined by sulphonation with sodium
disulphonate and sodium sulphate.

16. Disadvantage :-