NMY51 Yeast Strains

$255.00

  • Model: S0094
  • 49 Units in Stock
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NMY51 Yeast Strains

Alternative name      NMY51 Yeast Strains,NMY51,Yeast
Packing 100ul             Storage at -20centigrade/span>
Strains Resistance    Kan
Culture Medium       YPDA
Condition                28centigrade,under the aerobic conditions
Transformation       Electro Transformation

 

NMY51 Genotype

MATahis3Δ200trp1-901leu2-3,112ade2LYS2::(lexAop)4-HIS3ura3::(lexAop)8-lacZ ade2::(lexAop) 8-ADE2 GAL4

 

NMY51 Description

Dualmembrane system is a special screening technique developed by Dualsystems Biotech company to screen the interaction between transmembrane proteins, which uses separate ubiquitin system (split-ubiquitin) to directly detect the interaction between membrane proteins in natural state, is currently the only yeast double hybridization system on the market to detect the interaction between membrane proteins.

The system uses NMY51 yeast strain, which can directly transform the plasmid for protein mutual verification or screening test.


Transformation marker: TRP1, leu2-3

Reporting genes: HIS3, ADE2 and LacZ

 

NMY51 
1. Selective growth screening was carried out through the nutritional defect reporting gene (HIS3, ADE2),

2. Quantitative or semi-quantitative screening of beta-galactose analysis by LacZ reporting Gene, three independent reporting genes, regulated by different promoter, reduced the probability of false positives.

Principle: ubiquitin (ubiquitin) molecular weight is very small, composed of 76aa residues; ubiquitin, as a degradation signaling molecule, can be connected to the N end of another protein and then identified by ubiquitin specificity protease (ubps), resulting in enzymatic hydrolysis of proteins linked to ubiquitin. Ubiquitin can be artificially divided into two parts: the N end (Nub) and the C End (Cub). 

First, the 3-bit isoleucine of ubiquitin Nub was artificially mutated into glycine (Nubi mutation was NUBG). In this way, the affinity with Cub greatly reduced, avoiding the possibility of self-union or proximity of Cub and Nub. Secondly, the cub part is fused with the synthetic LEXA-VP16 transcription activating factor into a fusion protein cub-lexa-vp16. Under normal conditions, NUBG does not bind to Cub, ubps also does not recognize the separation of ubiquitin, transcription activation factor will not be cut off. Finally, the proteins to be tested are fused with NUBG and cub respectively to form bait fusion eggs (bait-cub-lexa-vp16) and Prey fusion proteins (PREY-NUBG).

If bait and prey interact, it will cause NUBG and Cub to approach each other and be ubps identified, leading to the dissociation of the lexa-vp16 and into the nucleus, thus activating the transcription of the reporting gene.

This system can use four kinds of bait plasmid: pbt3-n,pbt3-suc,pbt3-ste,pbt3-c, screening signs are leu; three prey plasmid: Ppr3-c, Ppr3-suc,ppr3-ste, screening signs are TRP.
The NMY51 yeast strain can be cultured YPDA under 28 centigradeaerobic conditions, and then the strain is preserved using 30% glycerol.

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