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[공식 대리점] Pronetbio-Yeast One Hybrid (Gene-Centered) Vector Kit, 효모 원-하이브리드 (유전자 중심) 벡터 키트,RY8009
- AI바이오허브 오래 전 2025.01.28 21:59 Bulletin Board 인기
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Yeast One Hybrid (Gene-Centered) Vector Kit
CAT：RY8009
Advanced Yeast One-Hybrid System for DNA-Protein Interaction Studies
The Yeast One Hybrid (Gene-Centered) Vector Kit is designed for researchers studying DNA-protein interactions to explore gene expression regulation, identify DNA binding sites, and analyze DNA-binding domains. Developed from the Yeast Two-Hybrid system, this kit offers a reliable, streamlined approach to uncover key interactions in eukaryotic cells.
Product Overview
This kit includes the Y187 strain (MATα type) and essential plasmids pHIS2 and pGADT7. The yeast one-hybrid assay leverages GAL4 AD to activate HIS3 gene expression when prey proteins bind to the bait DNA sequence. Growth on 3-AT supplemented media confirms DNA-protein interaction. The included primers, plasmids, and controls facilitate amplification, sequencing, and experimental validation.
Key Features:
Positive Control: pHIS2-p53 + pGAD53m;
Selection Markers: Trp for pHIS2, Leu for pGADT7;
Includes high-quality primers: pHIS2-F, pHIS2-R, T7-F, AD-R.
Product Components
Component | Name | Concentration | Specification | Quantity |
Plasmid | pGADT7 | 100 ng/μL | 20 μL | 1 tube |
Plasmid | pHIS2 | 100 ng/μL | 20 μL | 1 tube |
Plasmid | pHIS2-p53 | 100 ng/μL | 20 μL | 1 tube |
Plasmid | pGAD53m | 100 ng/μL | 20 μL | 1 tube |
Strain | Y187 | Inoculated Plate | Diameter: 90mm | 1 plate |
Primer | pHIS2-F | Lyophilized powder | 2 OD | 1 tube |
Primer | pHIS2-R | Lyophilized powder | 2 OD | 1 tube |
Primer | AD-R | Lyophilized powder | 2 OD | 1 tube |
Primer | T7-F | Lyophilized powder | 2 OD | 1 tube |
Storage and Shipping Conditions
Plasmids: Store at -20°C; shipped at room temperature.
Strains: Store and ship at room temperature.
Primers: Store at -20°C; shipped at room temperature.
Experimental Guidelines
Precautions
1. To ensure strain viability, streak the strain onto a YPDA agar plate before liquid culturing.
2. Store strains at -80°C with 50% glycerol to prevent repeated freeze-thaw cycles.
3. This product is for research use only and not for clinical, diagnostic, or therapeutic applications.
4. Maintain sterile conditions to avoid contamination.
FAQs
1. Why is self-activation testing required?
Self-activation is a common phenomenon where the promoter induces background expression of the reporter gene. Testing ensures that the bait DNA does not produce false positives.
2. Can non-transcription factor proteins be identified?
Yes. Yeast one-hybrid assays detect all proteins binding to the bait DNA, including non-transcription factors and potential false positives.
3. Why doesn't the Y187-pHIS2 system require staining reagents?
Unlike the EGY48-LacZ system, Y187-pHIS2 confirms interactions through HIS3 expression on selective media.
4. Is the mating method applicable for Y1H?
No. Both Y1HGold and Y187 strains are MATα type, and mating requires MATa and MATα strain compatibility.
Self-prepared Materials
1. Sterile PCR tubes, tips, centrifuge tubes, shakers, incubators, and reagents like 50% PEG3350, 1M LiAc, X-α-Gal, and 3-AT.
2. (Optional from Pronetbio) Yeast media (YPDA, SD-Trp, SD-Trp/-Leu/-His), yeast plasmid extraction kits, and positive clone amplification kits.
