Services
Providing Advanced Biological Technical Services for Researchers in Fundamental Science
Abiotic-Stress Resistance Gene Screening and Validation Services
Service Overview
ProNet Biotech offers advanced yeast-based high-throughput gene screening and validation services for identifying genes related to abiotic stress resistance. Our state-of-the-art system rapidly identifies genes from non-model stress-resistant species, helping researchers discover and validate genes involved in stress tolerance. Using yeast in a stress screening system, we perform gradient resistance experiments across the genome to identify resistance-related genes. Proteomics is then applied to identify candidate proteins, followed by phenotypic validation at the protein level.
High-Throughput Stress-Resistant Gene Library Construction
Our service includes the construction of high-throughput yeast gene libraries, screening under optimized stress conditions, and subsequent validation of candidate genes. With our technology, we ensure rapid identification of multiple stress-resistant genes, particularly for stressors like drought, salinity, and temperature extremes.
Library Construction Service Content and Deliverables
Library Construction Procedures (SMART method) | Working days | Deliverables |
1. RNA Extraction 2. mRNA Extraction | 5 | 1. 4 mL Yeast Library Glycerol Stock (divided into 4 tubes) (optional) 2. Yeast Library Plasmid (4 tubes > 400 µg) (optional) 3. Project Report in Word Document Format 4. Documentation Commitment to Meet the Following Criteria: Library Capacity﹥1*107 CFU, Average Insert Size 0.8-1.5 kb (varies by species), Empty Vector Rate < 5% |
3. cDNA Synthesis and Adaptor Ligation (Three-Frame Library) 4. cDNA Normolization | 5 | |
5. Recombinant Cloning into Yeast Vector pYES2-NTB and Electroporation 6. Electroporation of the cDNA Library into Competent E. coli Cells for Clone Amplification | 5 | |
| 7. Library Screening and Plasmid Extraction | 5 | |
| 8. Data Compilation and Analysis | 2 | |
9. Transcriptome Sequencing 10. Whole Library Sequencing 11. GO Analysis + KEGG Analysis | 15 |
High-Throughput Stress-Resistant Gene Screening
Gene Screening Service Workflow and Deliverables
High-Throughput Stress-Resistant Gene Screening | |||
Items | Experimental Procedures | Working days | Deliverables |
| Yeast Working Culture Preparation | 1. Yeast Working Culture Preparation 2. PCR Quality Control of Yeast Working Culture | 15 | 1. NGS Sequencing Results of Positive Clones (Dependent on Sequencing Results) 2. Sanger Sequencing Results 3. Project Report in Word Document Format (Including Experimental Data) 4. GO + KEGG Analysis Results |
| Optimization of Stress Conditions | 3. Prepare Media with Five Different Concentrations of Stress Conditions (Optimization of Stress Conditions) | 10 | |
| Yeast Screening | 4. Plate Yeast Working Culture: 20 Plates of 15 cm | 5 | |
| Identification of Positive Clones | 5. PCR Identification of Yeast Colonies 6. Sanger Sequencing | 10 | |
| Validation of Screening Results | 7. Validation of Screening Results 8. Data Compilation and Analysis | 10 | |
| NGS Sequencing | 9. NGS Sequencing 10. Data Compilation and Analysis | 15 | |
Stress-Resistant Gene Phenotypic Validation
Once candidate genes are identified through screening, phenotypic validation is performed to confirm their role in stress resistance:
Construct Overexpression Vector
The target gene is cloned into a yeast overexpression vector (pYES2).
Transform into Target Strain
The recombinant plasmid is transformed into the target yeast strain for expression and validation.
Control Transformation
An empty vector is transformed into the control strain for comparison.
Gradient Dilution Spot Plating
The transformed strains are plated under stress conditions for phenotypic verification.
Stress Treatment Medium
The strains are subjected to various stress treatments (high temperature, salinity, drought, etc.) to validate their stress resistance phenotype.
Stress resistance gene phenotype verification related strains | |
BY4741 | High temperature and low temperature resistance |
INVSC1 | Salt tolerance and drought tolerance |
∆Ycf1 | Heavy metal tolerance |
W303 | Pesticide resistance |
Phenotypic Validation Service Content and Deliverables
Items | Working days | Delivery material |
1.Construct target gene into pYES2 vector | 15 | 1. Target gene plasmid 2. Project Report in Word Document Format (Including Experimental Data) |
2. Stress condition experiment | 15 |
Technical Advantages
High-Throughput Screening
Our yeast-based system allows for the rapid screening of a large number of genes involved in abiotic stress resistance, enabling quick identification of stress-tolerant genes.
Soluble Eukaryotic Expression
Proteomics is used to identify candidate proteins for phenotypic validation at the protein level, ensuring accurate validation of gene functions.
Sensitive Yeast Strains
Yeast strains like YCF1, BY4741, and InVSCl are unicellular eukaryotic microorganisms that grow rapidly, are easy to manipulate genetically, and can secrete expressed proteins. These strains also perform post-translational modifications, making them ideal for expressing foreign genes.
Why Choose ProNet Biotech?
At ProNet Biotech, we leverage the latest in high-throughput yeast screening and validation technologies to provide you with accurate, fast, and cost-effective solutions for your research. Whether you are working on stress-tolerant crop development or environmental stress-related gene discovery, our comprehensive services are designed to meet your needs.
Ready to Get Started?
Contact us below to learn more about how our yeast-based abiotic-stress resistance gene screening and validation services can accelerate your research. We look forward to partnering with you in discovering the next breakthrough in stress tolerance!
Disclaimer: Our services are sold for research purposes only and are not intended for diagnostic or therapeutic use!
Services Workflow
Online Consultation
01
Solution Matching
02
Service Contract
03
One-Stop-Services
04
Project Report
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Contact Us
Tel: +86 025-85205672
Email: info@pronetbio.com
Address: Building 3C, Nanjing Xianlin Zhigu,
Qixia District, Nanjing, Jiangsu, China, 210033
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