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Recombinant Protein and
Its Expression Systems
What is
Recombinant
Protein ?
What does Proteins do?
This is what we have in nature
This is the quantity and
purity we want
Why We Need Recombinant Protein
“Recombinant protein is a manipulated form of protein, which is
generated in various ways to produce large quantities of p...
Ori
pCDNA3.1
5428bp
Amp r
Neomycin
SV40 ori
PCMV
f1 ori
BGHpA
SV40 pA
T7
Recombinant DNA/
Vector
Host Cell+ Vector
Host Ce...
Choose Suitable Expression System
Target Protein Property
Application
Protein Yield
Cost
Post-translational Modificat...
Choose Suitable Expression System
Bacteria
Expression
System
Yeast
Expression
System
Insect
Expression
System
Mammalian
Ex...
Bacteria Expression System
Advantage:
Clear genetic background
Fast breeding
Low cost
High expression level
Easy puri...
Bacteria Expression System
Disadvantage:
Hard to purified inclusion bodies
Inclusion Bodies
Aggregation
Bacteria Expression System
Disadvantage:
Imperfect post-
translational modification
processing system
Low biological act...
Yeast Expression System
Sequenced in 1996 Used in brewing and bread industry for
thousands of years
Yeast Expression System
Disadvantage of
S.cerevisiae :
Not suitable for high-
density culture
Lacking strong and strict
...
Yeast Expression System
Methanol Yeast
System Host Cells:
H Polymorpha
Candida Bodini
Pichia Pastoris
Fission Yeast Met...
Yeast Expression System
Yeast Expression System
Nucleus
Cytoplasm
Mxr1
Mxr1
Glucose
Methanol Prm1 Mit1 Aox1
Advantage:
Production up to grams
Tr...
Baculovirus Expression System
Generation of
Recombinant Baculovirus
and
Gene Expression
Limitation Factors:
Culture mediu...
Baculovirus Expression System
Baculovirus Expression System
Advantages:
Recombinant protein has complete biological function
Post-translation modifica...
Mammalian Expression System
Plasmid
Transfection
Viral Vector
Infection
Mammalian Expression System
Transient Expression
System:
Short Protein
Expression Duration
Simple
Short Time
Stable Cel...
Mammalian Expression System
Advantages:
 Initiation Signals
Protein Processing
Protein Secretion
Protein Glycosylation...
Bacterial
Expression
System
Yeast Expression System
Insect Expression
System
Mammalian Expression
System
Speed ★★★★ ★★★ ★★...
Contact Information:
Phone: 1-631-559-9269
1-631-448-7888
Fax: 1-631-938-8127
Address: 45-1 Ramsey Road, Shirley,
NY 11967...
Thanks!
Any questions?
You can find me at contact@creative-biomart.com
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The video explain what is recombinant protein and recombinant expression systems including bacterial system, yeast system, insect system and mammalian system. And how to choose suitable expression system.

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Recombinant Protein and Its Expression Systems

  1. 1. Recombinant Protein and Its Expression Systems
  2. 2. What is Recombinant Protein ?
  3. 3. What does Proteins do?
  4. 4. This is what we have in nature This is the quantity and purity we want Why We Need Recombinant Protein
  5. 5. “Recombinant protein is a manipulated form of protein, which is generated in various ways to produce large quantities of proteins, modify gene sequences and manufacture useful commercial products. Recombinant protein is encoded by recombinant DNA, which has been cloned in a system that supports expression of the gene and translation of mRNA.
  6. 6. Ori pCDNA3.1 5428bp Amp r Neomycin SV40 ori PCMV f1 ori BGHpA SV40 pA T7 Recombinant DNA/ Vector Host Cell+ Vector Host Cell Proliferation High-level Protein Expression Recombinant Protein Technology
  7. 7. Choose Suitable Expression System Target Protein Property Application Protein Yield Cost Post-translational Modifications …
  8. 8. Choose Suitable Expression System Bacteria Expression System Yeast Expression System Insect Expression System Mammalian Expression System
  9. 9. Bacteria Expression System Advantage: Clear genetic background Fast breeding Low cost High expression level Easy purification Good stability Strong anti-pollution ability Wide application range
  10. 10. Bacteria Expression System Disadvantage: Hard to purified inclusion bodies Inclusion Bodies Aggregation
  11. 11. Bacteria Expression System Disadvantage: Imperfect post- translational modification processing system Low biological activity
  12. 12. Yeast Expression System Sequenced in 1996 Used in brewing and bread industry for thousands of years
  13. 13. Yeast Expression System Disadvantage of S.cerevisiae : Not suitable for high- density culture Lacking strong and strict regulation promoter Low secretion efficiency
  14. 14. Yeast Expression System Methanol Yeast System Host Cells: H Polymorpha Candida Bodini Pichia Pastoris Fission Yeast Methanol Yeast
  15. 15. Yeast Expression System
  16. 16. Yeast Expression System Nucleus Cytoplasm Mxr1 Mxr1 Glucose Methanol Prm1 Mit1 Aox1 Advantage: Production up to grams Translation closer to mammalian cells Don’t undergo hyperglycosylation
  17. 17. Baculovirus Expression System Generation of Recombinant Baculovirus and Gene Expression Limitation Factors: Culture medium Oxygen supply Logarithmic growth …
  18. 18. Baculovirus Expression System
  19. 19. Baculovirus Expression System Advantages: Recombinant protein has complete biological function Post-translation modification High level of expression, up to 50% of the total protein amount Accommodate large insert protein Simultaneously express multiple genes Safer to use than mammalian virus …
  20. 20. Mammalian Expression System Plasmid Transfection Viral Vector Infection
  21. 21. Mammalian Expression System Transient Expression System: Short Protein Expression Duration Simple Short Time Stable Cell Line Generation: Time-consuming and laborious Protein Expression Last a Long Time
  22. 22. Mammalian Expression System Advantages:  Initiation Signals Protein Processing Protein Secretion Protein Glycosylation Intact Macromolecules Native Structure … Disadvantages: Complicated High Requirement Low Yield Sometimes Viral Infections …
  23. 23. Bacterial Expression System Yeast Expression System Insect Expression System Mammalian Expression System Speed ★★★★ ★★★ ★★ ★ Yield ★★★ ★★★★ ★★ ★ PTM (relative to human) ★ ★★ ★★★ ★★★★ Cost ★★★★ ★★★ ★★ ★ Application Prokaryotic protein, simple eukaryotic protein Intracellular/Secreted Protein, Disulfide-bonded protein, Glycosylated protein Membrane protein, Large-size protein, Viral vaccines, signaling proteins, cytokines, kinases Complex eukaryotic protein, Protein need accurate PTM Simply Compare among Expression Systems
  24. 24. Contact Information: Phone: 1-631-559-9269 1-631-448-7888 Fax: 1-631-938-8127 Address: 45-1 Ramsey Road, Shirley, NY 11967, USA Email: info@creative-biomart.com
  25. 25. Thanks! Any questions? You can find me at contact@creative-biomart.com
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The video explain what is recombinant protein and recombinant expression systems including bacterial system, yeast system, insect system and mammalian system. And how to choose suitable expression system.

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