Yeast two-hybrid Technical Service丨Zoonbio

Yeast two-hybrid Technical Service

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Summary

Yeast two-hybrid can reflect the interaction of different proteins in living cells. It has been widely used in the study of gene function with the characteristics of simplicity, sensitivity and efficiency.

Zoonbio can provide yeast two-hybrid experimental services for cytoplasmic protein, nucleoprotein and membrane protein. We have rich experience in yeast two-hybrid technical services, which can help customers analyze and solve various problems encountered in the experimental process.

Service Items

service items	explain	cycle
technical services for construction of yeast single hybrid library	library construction scheme based on reorganization method	4-6 weeks
screening technology service of yeast single hybrid library	DNA-protein interaction	12-15 weeks
screening of yeast two-hybrid libraries in nucleoprotein system	based on pDEST22 or pGADT7 system	14-16 weeks
screening of yeast two-hybrid library in Membrane protein system	separated ubiquitin-mediated yeast two-hybrid system for membrane protein	14-16 weeks

case

1.construction and identification of bait vector pGBKT7-ORF1. 2.ORF1 self-activation detection

All transformants can grow normally on SD-TL plate; on SD-TLHA defective plate, positive control transformants could grow, but pGBKT7-ORF1+pGADT7 transformants could not grow, which was the same as negative control; the results of transformant pGBKT7-ORF1+pGADT7 in LacZ assay were the same as those in negative control, so ORF1 bait cloning had no self-activating effect.
3.library transformation
4.detection of reporter genes of positive clones of His, Ade and LacZ.

Mark "+" as positive control,"-" as a negative control. Thirteen positive strains containing Histidine and Adenine genes could grow on SD-HLHA plate. Twelve of the 13 positive strains screened initially could activate LacZ gene, of which 12, 16 and 17 strains were if activated.
5.extraction and sequencing comparison of yeast positive clonal DNA
6.rotary verification

Mark "+" as positive control,"-" as a negative control. All six positive strains could grow on SD-THLA plate and activate LacZ gene.