Services
- Yeast Hybrid Services
- Yeast One-Hybrid (Y1H) System Service
- Yeast One-Hybrid (Y1H) Library Construction Service
- Yeast One-Hybrid (Y1H) Library Screening Service
- Prokaryotic Yeast One-Hybrid (Y1H) Service
- Genomic Yeast One-Hybrid (Y1H) Service
- Yeast One-Hybrid (Y1H) Point-to-Point Verification Service
- Yeast Library Construction Service for Transcription Factors
- Yeast Library Screening Service for Transcription Factors
- Transcription Factor Self-Activation Verification Service
- Yeast Two-Hybrid (Y2H) System Service
- Nuclear System Y2H Library Construction Service with Invitrogen Gateway Method
- Nuclear System Y2H Library Construction Service with Clontech-SMART Method
- Membrane System Y2H Library Construction Service with Invitrogen-Gateway Method
- Membrane System Y2H Library Construction Service with Clontech-SMART Method
- Reverse Yeast Two-Hybrid (rY2H) Library Construction Service
- Yeast Two-Hybrid (Y2H) Library Screening Service
- Nuclear System Yeast Two-Hybrid (Y2H) Library Screening Service
- Membrane System Yeast Two-Hybrid (Y2H) Library Screening Service
- Nuclear System Yeast Two-Hybrid (Y2H) Verification Service
- Membrane System Yeast Two-Hybrid (Y2H) Verification Service
- High-throughput Yeast Two-Hybrid (Y2H) Library Screening Service
- Prokaryotic Yeast Two-Hybrid (Y2H) Service
- Genomic Yeast Two-Hybrid (Y2H) Service
- Mammalian Two-Hybrid (mY2H) Service
- Yeast One-to-One Verification Service
- Yeast Three-Hybrid (Y3H) System Service
- Yeast One-Hybrid (Y1H) System Service
- Yeast Expression System Services
- Yeast Foreign Gene Secretion and Expression Service
- Yeast Signal Peptide Tracking and Screening Service
- Yeast Stress Resistance Gene Screening Service
- Yeast Gene Knockout Service
- Yeast Surface Display Service
- Yeast Display Service of IgG Full-length Antibody
- Pichia pastoris Protein Expression Analysis
- Saccharomyces cerevisiae Protein Expression Analysis
- DNA Shuffling
- Yeast RNAi System
- Yeast Detection Services for Protein Functional Domain
- Yeast Microarray Chip Service
Online Inquiry
Pichia pastoris Protein Expression Analysis
Yeast expression system has the advantages of both prokaryotic and advanced eukaryotic systems. It has the characteristics of ordinary culture conditions, fast growth, relatively low cost, capable of post-translational protein processing, and easy access to soluble active recombinant proteins. It is widely used in the production of recombinant proteins, especially eukaryotic proteins. Among them, Pichia pastoris has the advantages of fast growth, abundant commercial expression vectors, and high-efficiency secretion expression, making it the preferred choice for yeast expression of recombinant proteins.
The Pichia pastoris expression system is a eukaryotic expression system that has developed rapidly and is widely used in recent years. It is a methanol-trophic yeast with two alcohol oxidase (AOX) encoding genes, AOX1 and AOX2. The two sequences are similar. The AOX1 gene is strictly induced and regulated by methanol. When methanol is the sole carbon source, the AOX1 promoter can be induced by methanol to initiate the expression of ethanol oxidase, thereby metabolizing with methanol. The plasmid containing the AOX1 promoter can be used to promote the expression of the target gene encoding the foreign protein.
Service introduction
The yeast protein expression platform of Creative BioMart is composed of experimental staff who have been engaged in yeast expression operations for many years. They have rich experience and successfully expressed more than 100 kinds of recombinant proteins, which can achieve the highest quality protein expression to the greatest extent, and can meet a variety of special needs of customers.
Characteristics of Pichia pastoris expression strains
- GS115
- KM71
Both AOX1 and AOX2 genes normally express AOX protein (alcohol oxidase), the utilization rate of methanol is high, close to the wild type, and belongs to the rapid utilization type of methanol (Mut+)
The AOX1 gene is mostly deleted, and only AOX2 can be used to encode AOX. The utilization rate of methanol slows down, showing the slow utilization of methanol (Muts).
| Restriction enzyme | Insertion site | GS115 phenotype | KM71 phenotype |
|---|---|---|---|
| Sa1l / Stul | HIS4 | HIS4 Mut+ | HIS + MutS |
| Sacl | 5' AOX1 | HIS + Mut+ | HIS + MutS |
| Bg1lI | Replace AOX1 | HIS + MutS | HIS + MutS |
Commonly used expression vectors and selection markers
Vector information
| Expression type | Vector name | Selection marker | Promoter |
|---|---|---|---|
| Intracellular expression | pA0815 | HIS4 | AOX1 |
| pPIC3K | HIS4 | AOX1 | |
| pPUC3.5K | HIS4 | AOX1 | |
| Secretory expression | pPICZa, A, B, C | Bler | AOX1 |
| pPIC9K | HIS4 | AOX1 |
Signal peptide
For extracellularly expressed proteins, in addition to the signal peptide that comes with the vector, the following signal peptides are available for selection.
Signal peptide sequence
| Signal peptide | Protein sequence |
|---|---|
| αMF* | MRFPS IFTAV LFAAS SAL AA PVNTT TEDET AQIPA EAVIG YSDLE GDFDV AVL PF SNSTN NGLLF INTTI ASIAA KEEGV SLEKR EAEA |
| Dse4 | MSFSS NVPQL FLLLV LLTNI VSG |
| Exg1 | MNLYL ITLLF ASLCS A |
| Scw11 | MLSTI LNIFI LLLFI QASLQ |
| Salb | MKWVT FISLL FLFSS AYS |
Service items
| Service items | Service cycle (weeks) |
|---|---|
| Protein analysis and codon optimization | Finish before the project starts |
| Gene synthesis | 2~4 |
| Subcloning | 1 |
| Gene expression purification test | 2 |
| Fermentation and purification | 1~2 |
| total | 6~9 |
Experiment process
Service process
Please let us know the details of your yeast related projects through online consultation. We will reply to you in time and provide you with a comprehensive evaluation report on your project, including the feasibility evaluation of the project and the detailed experimental plan designed by our professionals. As our condition optimization is completed, the result report and other final products will be delivered to you immediately.
Creative BioMart has accumulated many years of experience in the field of yeast research, helping our customers accelerate research in all fields using yeast as a research tool, and improve the overall success rate of the project. Please tell us your project requirements, and we will provide you with a full service from strategy design to final report. If you have any questions, please feel free to contact us.