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Yeast cells Expression
Yeast protein expression system is the most economical eukaryotic expression system for both secretion and intracellular expression. It is ideally suitable for large-scale production of recombinant eukaryotic proteins.
GeneCreate Technology, selected stable and durable production strains that resemble mammalian system are employed for high-yield, high-productivity protein processing, thus dramatically reducing the associated costs and inherent time requirements. Otherwise, our Yeast protein expression systems serve as a eukaryotic expression system integrate the advantages of the mammalian cell expression system.So the protein expressed by yeast protein expression system could be modificated such as glycosylation, acylation, Phosphorylation and so on to ensure that the protein native conformation.So It could Used to produce protein material with high added value that is very close to natural protein. The facts proved that our protein produced by Yeast protein expression system which served as raw materials for downstream preparation monoclonal antibody had good properties of the recognition of natural protein through the feedback of the customs.
Combining advanced technologies
and profound past experience, our efficient seed screening work can make your
order to obtain the highest yield in the shortest time.
Advantages
l Utilizing Biobrick technology
patents combine with our reformed efficient secretion vector ,thus Can
construct any copies of the gene dose in vitro with efficiency, enhance the expression quantity,
and could be successfully used in the optimization of improving industrial
strains, makes the maximum protein expression.
2 GeneCreate can take your project directly from gene synthesis, to downstream activities including recombinant protein expression, stable cell line generation, antibody production, and assay development,more worthy of mention is our homogeneous gene mutant library, is a global innovation technology.
Service menu
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Yeast protein expression system economical menu service |
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Milestones |
Specifications |
Deliverables |
Timeline |
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1. protokaryon cloning |
Genes obtain |
Nucleic acid electrophoresis pictures |
2 weeks |
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PCR amplification product connected to Ppic9k modified by our company Transformate to DH5 α |
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Large-scale synthesis of recombinant plasmid |
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Linearize the recombinant plasmid |
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2. Expression optimization |
Electric Transformation of the appropriate yeast host strain |
Nucleic acid electrophoresis pictures |
2-3 weeks |
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PCR verification of 10 strains randomly selected from transformation plate |
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3. Protein expression and purification |
randomly select a PCR verification positive strain for 1 L large-scale culture |
Expressed supernatant |
1-2 weeks |
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The dispose of the supernatant and One-step purification |
Purified product SDS-PAGE and western blot |
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Purified product SDS-PAGE and western blot |
1L supernatant Purified product |
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Yeast protein expression system service standard menu |
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Milestones |
Specifications |
Deliverables |
Timeline |
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1. protokaryon cloning |
Genes obtain |
Nucleic acid electrophoresis pictures |
2 weeks |
|
PCR amplification product connected to Ppic9k modified by our company，Transformate to DH5 α |
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Linearize the recombinant plasmid |
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Linearize the recombinant plasmid |
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2. Expression optimization |
Electric Transformation of the appropriate yeast host strain |
Nucleic acid electrophoresis pictures |
5-6 weeks |
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The poly copies screening via genetic G418 |
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PCR identification of multi-copy cloning |
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Select more than 40 multi-copies strains for small-scale expression |
Purified product SDS-PAGE and western blot |
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3. Protein expression and purification |
Select the best strain for large-scale expression according to the customer request |
Expressed supernatant SDS-PAGE < |
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Example
Analyse
GeneCreate has successfully applied the yeast expression system for producing various proteins:
A B
Fig. A: Expression and secretion of target protein from P.pastoris.
Fig.B: Purification of the target protein by Ni column.
C D
Fig. C: Expression and secretion of pharmaceutical protein from P.pastoris.
Lane 1 :Expression level from customer.
Lane 2 :Expression level from CUSABIO.
Lane 3 :Negative control.
Fig.D: Purification of the target pharmaceutical protein by SP-Sepharose.
Fig. E: single copy gene expression of industrial enzyme from P.pastoris.
Fig.F: eight copy gene expression of industrial enzyme from P.pastoris.
Remarks： We only provide HIS tag antibody used to the quality control of expression product, if customer's requires do western blot with a corresponding protein antibodies, the antibodies are provided by the customers themselves.