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Genetic engineering
Recombinant DNA technology
Bacterial plasmids
•
Most bacterial DNA is
on a single large
chromosome, but some
DNA is in a small circle
called a plasmid.
Bacterial Plasmids in Nature
1-Occur naturally in bacteria and usually
carry genes that are useful but not
essential to survival: e.g. genes which
make bacteria resistant to antibiotics.
2-Plasmids are released by dead bacteria
and absorbed by those still living thus
genetic information is exchanged (sexual
reproduction?).
Bacterial Plasmids in Nature
3-Some plasmids even contain
genes that build a transfer tube
between bacteria.
4-There can be as many as
several hundred copies of a
single plasmid in each bacteria.
FUNCTIONS OF PLASMIDS
1-the main function is to carry antibiotic resistance genes
2-carry genes involved in metabolic activities and digesting pollutants
3-producing antibacterial proteins
4-carry genes which are concerned with increasing pathogenicity of
bacteria
5- using in cloning process in humans
Classifications of plasmids
1-conjugative plasmids :
Conjugative plasmids contain a set of transfer or
tra genes which promote sexual conjugation
between different cells
2-non-conjugative plasmids:
Non-conjugative plasmids are incapable of
initiating conjugation, hence they can be
transferred only with the assistance of
conjugative plasmids.
TYPES OF PLASMIDS ACCORDING TO FUNCTIONS
1-Fertility F-plasmids, which contain transfer or tra genes. They are capable
of conjugation and result in the expression of sex pilli.
2-Resistance (R) plasmids, which contain genes that provide resistance
against antibiotics or poisons. Historically known as R-factors, before the
nature of plasmids was understood.
3-Col plasmids, which contain genes that code for bacteriocins, proteins that
can kill other bacteria.
4-Degradative plasmids, which enable the digestion of unusual substances,
e.g. toluene and salicylic acid.
5- Virulence plasmids, which turn the bacterium into a pathogen
Plasmid Structure
1- Plasmids only need an “origin or
replication” and a “useful” gene to be
considered complete.
2-Molecular biologists have been able to
“insert” custom built restriction sites into
many plasmids so they can be used to
“insert” DNA fragments from other genes
into them and thus have a way to
propagate those DNA pieces.
Plasmids can be manipulated easily in the laboratory
1-Plasmids can be collected from bacteria.
2-Restriction enzymes can be purified and used to identify and cut out specific
sequences of DNA along with the plasmid vector.
3-Ligase (enzyme) can be purified and used to “glue” pieces of DNA together.
4-Bacteria can be transformed by taking in plasmids given to them
How to produce human insulin from bacteria and become a
multimillionaire
1-Isolate plasmid DNA from bacteria and
insulin gene from human.
2-Cut both DNAs with the same
restriction enzyme.
3-Mix the DNA together with ligase.
4-Insert the new DNA into bacteria
(transformation).
5-Use antibiotics to kill any bacteria
without the plasmid.
6-Grow bacteria and harvest the insulin.
Insulin
Purification
Insulin gene
Human DNA
Thank you

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plasmid .genetic engineering.pptx

  • 2. Bacterial plasmids • Most bacterial DNA is on a single large chromosome, but some DNA is in a small circle called a plasmid.
  • 3. Bacterial Plasmids in Nature 1-Occur naturally in bacteria and usually carry genes that are useful but not essential to survival: e.g. genes which make bacteria resistant to antibiotics. 2-Plasmids are released by dead bacteria and absorbed by those still living thus genetic information is exchanged (sexual reproduction?).
  • 4. Bacterial Plasmids in Nature 3-Some plasmids even contain genes that build a transfer tube between bacteria. 4-There can be as many as several hundred copies of a single plasmid in each bacteria.
  • 5. FUNCTIONS OF PLASMIDS 1-the main function is to carry antibiotic resistance genes 2-carry genes involved in metabolic activities and digesting pollutants 3-producing antibacterial proteins 4-carry genes which are concerned with increasing pathogenicity of bacteria 5- using in cloning process in humans
  • 6. Classifications of plasmids 1-conjugative plasmids : Conjugative plasmids contain a set of transfer or tra genes which promote sexual conjugation between different cells 2-non-conjugative plasmids: Non-conjugative plasmids are incapable of initiating conjugation, hence they can be transferred only with the assistance of conjugative plasmids.
  • 7. TYPES OF PLASMIDS ACCORDING TO FUNCTIONS 1-Fertility F-plasmids, which contain transfer or tra genes. They are capable of conjugation and result in the expression of sex pilli. 2-Resistance (R) plasmids, which contain genes that provide resistance against antibiotics or poisons. Historically known as R-factors, before the nature of plasmids was understood. 3-Col plasmids, which contain genes that code for bacteriocins, proteins that can kill other bacteria. 4-Degradative plasmids, which enable the digestion of unusual substances, e.g. toluene and salicylic acid. 5- Virulence plasmids, which turn the bacterium into a pathogen
  • 8. Plasmid Structure 1- Plasmids only need an “origin or replication” and a “useful” gene to be considered complete. 2-Molecular biologists have been able to “insert” custom built restriction sites into many plasmids so they can be used to “insert” DNA fragments from other genes into them and thus have a way to propagate those DNA pieces.
  • 9. Plasmids can be manipulated easily in the laboratory 1-Plasmids can be collected from bacteria. 2-Restriction enzymes can be purified and used to identify and cut out specific sequences of DNA along with the plasmid vector. 3-Ligase (enzyme) can be purified and used to “glue” pieces of DNA together. 4-Bacteria can be transformed by taking in plasmids given to them
  • 10. How to produce human insulin from bacteria and become a multimillionaire 1-Isolate plasmid DNA from bacteria and insulin gene from human. 2-Cut both DNAs with the same restriction enzyme. 3-Mix the DNA together with ligase. 4-Insert the new DNA into bacteria (transformation). 5-Use antibiotics to kill any bacteria without the plasmid. 6-Grow bacteria and harvest the insulin. Insulin Purification Insulin gene Human DNA