Services
Providing Advanced Biological Technical Services for Researchers in Fundamental Science
Ion Transport - Yeast Single-Gene Verification
Technical Principle
This service utilizes a genetically well-defined yeast (Saccharomyces cerevisiae) system to rapidly and economically verify the functions of target genes related to metal ion tolerance or homeostasis. The core method is based on functional complementation experiments conducted in a yeast mutant strain that is highly sensitive to metal ion overload or has inherent defects in ion absorption. By observing whether the introduced exogenous gene can enable the defective yeast mutant to grow under high/low ion concentration conditions, the function of the gene can be determined.
Application Scenario
1. Iron ions:
(1) Cloning and functional identification of genes related to iron absorption tolerance in plants
(2) Research on Genes Related to Iron Absorption and Transport in Plants
2. Zinc ions:
(1) Verify whether candidate genes from plants or animals (such as metal transport proteins, chelation peptide synthases, and transcription factors) are directly involved in zinc tolerance or homeostasis regulation
(2) Rapidly compare the functional differences among different members of the homologous gene family
(3) Conduct a preliminary mutation analysis on the identified genes and locate the functional key regions.
3. Potassium ions:
(1) Cloning and functional identification of genes for efficient low-potassium tolerance in plants
(2) Research on the Structural and Functional Relationship of Potassium Ion Channels/Transport Proteins
4. Cadmium ions:
(1) Mining and functional identification of cadmium ion transport genes in plants
(2) Research on the Regulatory Mechanism and Interactions of Cadmium Transport Genes
(3) Cadmium Pollution Control and Breeding of Cadmium-Tolerant Crops
Technical Advantages
1. Strictly set up blank controls: to reduce false positive rates and ensure the accuracy of the experiment.
2. Phenotypic comparison is intuitive: Through a simple plate seeding experiment, on different ion concentration gradient media, the phenotypic results of the target gene can be clearly observed at a glance.
3. Short experimental period: Compared to the transgenic validation conducted in plants (lasting from several months to several years), yeast validation can complete the entire process from transformation to phenotypic observation within 1-2 weeks, significantly saving time and resources.
4. Applicable for initial mechanism exploration: It enables the convenient construction of mutant forms of the target gene (such as point mutations, domain deletions), and allows for rapid testing of functional changes in defective yeast mutants, thereby preliminarily analyzing key functional sites.
5. Clear genetic background:
(1)CCC1 is an iron transporter protein in yeast. It is located on the vacuolar membrane and transports excess iron ions into the vacuole to maintain the iron balance within the yeast cell. Its dysfunction makes the yeast sensitive to excessive iron.
(2) Zinc-sensitive yeast strain zrc1 gene deletion mutant. This gene encodes a vacuolar membrane zinc transporter, which is responsible for isolating excess zinc ions into the vacuole to detoxify them. The zrc1 mutant is highly sensitive to ZnSO₄ in the environment, providing an ideal and specific genetic background for verifying whether the candidate gene can restore zinc tolerance.
(3) R5421 is a double-knockout mutant of Δ trk1Δtrk2, with extremely weak potassium absorption ability on its own. This eliminates the interference from endogenous potassium transport proteins in yeast.
(4) YCF1 is a gene encoding a vacuole membrane transporter in Saccharomyces cerevisiae. Its core function is to detoxify heavy metals, particularly playing a significant role in isolating and transporting heavy metals such as cadmium and chromium. Δycf1 is suitable for single-gene functional verification related to heavy metal detoxification/translocation, and is a classic model strain for single-gene verification in this field.
Supporting strains for verifying ion transport function | |
| Δccc1 | Iron transport defect type |
| Δzrc1 | Zinc transport defect type |
| R5421 | Potassium transport defect type |
| Δycf1 | Cadmium transport defect type |
Sampling Standards
| Type | Sample Submission Standards | Conditions Of Carriage | |
| Choose one from the three options | The sequence to be verified | Our company is engaged in gene synthesis. | |
| The constructed gene plasmid | Plasmid concentration ≥ 100 ng/μL; Plasmid volume ≥ 30 μL; There is a verification sequencing file | Ice bag | |
| Genetic plasmid bacterial solution | Each tube volume is ≥ 500 μL; There are identification and sequencing files | Dry ice delivery: Dry ice usage: 1 day (5 kg), 2-3 days (10-20 kg), 3-5 days (20-30 kg). | |
Cycle And Delivery
| Service content | Weekday (working day) | Delivery Content |
| The target gene was inserted into the pYES2-NTB vector | 10 | 1. The constructed recombinant plasmid 2. One electronic version of the experimental report |
| Minor adjustment assessment | 4 | |
| Yeast phenotype verification | 11 |
Note: Repeated point board installation requires additional steps and charges.
Case Show
Figure: pYES2-NTB-A enables the yeast strain Δzrc1 to normally transport Zn2+
Figure: pYES2-NTB-A enables the yeast strain ccc1 to normally transport Fe3+
Services Workflow
Online Consultation
01
Solution Matching
02
Service Contract
03
One-Stop-Services
04
Project Report
05
Related Products
Product Inquiry
Contact Us
Tel: +86 025-85205672
Email: info@pronetbio.com
Address: Building 3C, Nanjing Xianlin Zhigu,
Qixia District, Nanjing, Jiangsu, China, 210033
Need more info?
Let's connect!