Genetic Transformation of Bacteria: Methods and Applications in Biological Research and Pharmaceutical Production it was my presentation for lesson "research methods in medicine" with informations from Chatgpt

1. Genetic Transformation of Bacteria: Methods and
Applications in Biological Research and
Pharmaceutical Production
Zamanfar mahdi 118a

2. Introduction
we'll explore the fascinating world of genetic
transformation, focusing on its methods and
applications in scientific research and
pharmaceutical production. Our key element in this
journey is Calcium Chloride, a critical component in
one of the transformation methods.

3. Overview
Genetic transformation, in essence, is the process of introducing
foreign DNA into an organism's genome. In this presentation, we'll
delve into how this process is employed for scientific purposes.
Genetic transformation is pivotal in biotechnology, enabling us to
modify organisms for various purposes.
In particular, we'll discuss its crucial role in scientific research and
the pharmaceutical industry.

4. Genetic Transformation in Bacteria
Genetic transformation refers to the alteration of a
bacterium's genotype by the introduction of external
DNA. This can occur naturally or artificially.
For scientific purposes, we're primarily interested in
artificial transformation, where we control and direct
the genetic changes.
Understanding genetic transformation's role in
scientific research is essential for harnessing its
potential.

5. Methods of Genetic Transformation
Genetic transformation can be achieved through several methods, each with its advantages and applications.
Key methods include:
Chemical Transformation (Calcium Chloride)
Electroporation
Conjugation
Liposome-Mediated Transformation

6. Chemical Transformation with Calcium Chloride
Calcium Chloride plays a crucial role in chemical
transformation.
This method involves preparing competent cells, treating
them with calcium chloride, subjecting them to a heat
shock, and finally plating and selecting transformed
bacteria.
It's known for its high efficiency and cost-effectiveness.

7. Calcium Chloride Method
• Treatment with calcium
chloride in the early log
phase of growth for
Competence
• Bacterial cell membrane is
permeable to chloride ions,
but is non-permeable to
calcium ions
Chloride
Ions
E.Coli

8. Calcium Chloride Method
• As the chloride ions enter
the cell, water molecules
accompany the charged
particle
• Influx of water causes the
cells to swell and is
necessary for the uptake
of DNA
• The exact mechanism of
this uptake is unknown.

9. Calcium Chloride Method
• DNA of interest is then added to the
cells

10. Calcium Chloride Method
• Calcium chloride
treatment be followed
by addition of DNA of
interest then by heat.
• The heat shock step is
necessary for the
uptake of DNA.

11. Calcium Chloride Method
• Temperatures > 42degC: Bacteria’s
ability to uptake DNA reduces
• Extreme temperatures: Bacteria dies.

12. Calcium Chloride Method
• After the heat shock step intact plasmid
DNA molecules replicate in bacterial
host cells
• To help the bacterial cells recover from
the heat shock cells are briefly
incubated with non-selective growth
media

13. Calcium Chloride Method
• As the cells recover,
plasmid genes are
expressed
• Bacterial colonies
selected using antibiotic
selection techniques

15. Electroporation
Electroporation is another method for genetic transformation,
involving the use of electrical pulses to create temporary pores
in cell membranes.
It has its applications in situations where high efficiency is
required.

17. Conjugation
•Conjugation is a natural method of genetic transfer among
bacteria, where a donor bacterium transfers genetic material to
a recipient.
•It's essential for understanding horizontal gene transfer.

18. Liposome-Mediated
Transformation
•Liposome-mediated transformation
involves the use of liposomes to
transport DNA into cells.
•This method is useful for
specialized applications that require
precise control.

19. Applications in Scientific Research
•Genetic transformation in bacteria is invaluable for studying
gene functions, producing recombinant proteins, investigating
antibiotic resistance, and understanding pathogenicity.

20. Applications in Pharmaceutical Production
•In the pharmaceutical industry, genetic transformation of
bacteria is crucial for producing recombinant pharmaceuticals,
developing vaccines, enhancing antibiotic production, and
ensuring the quality of biopharmaceuticals.

21. Case Study: Insulin Production
•Let's examine a specific case study: the
production of insulin. Genetic transformation
of bacteria is used to produce insulin,
revolutionizing diabetes treatment.

22. Conclusion
•In conclusion, genetic transformation of bacteria is a
cornerstone of modern biotechnology.
•Understanding its methods and applications is essential for
advancing scientific research and pharmaceutical production.
•Calcium Chloride, as we've seen, plays a vital role in this
process.