The yeast two-hybrid assay was carried out using the Matchmaker gold yeast two-hybrid system (Clontech), according to the manufacturer’s instructions. Briefly, human Golgin45 (CC#1-3) and its mutants were subcloned in the GAL4 activation domain vector pGADT7 (Prey). The GTP or GDP-locked Rab2 mutants (Rab2-Q65L and Rab2-S20N) were subcloned into the Gal4 DNA binding domain vector, pGBKT7 (Bait). The Y2HGold yeast strains co-transformed with bait/prey pair were spreaded onto selective medium lacking leucine and tryptophan (SD/-Leu/-Trp, DDO, Clontech). The physical binding of bait and prey was identified by colony selection in selective medium lacking adenine, leucine, tryptophan and histidine (SD/-Ade/-Leu/-Trp/-His, Clontech) supplemented with X-α-Gal and Aureobasidin A (QDO/ X-α-Gal/AbA).
Qdo x α gal aba
Manufactured by Takara Bio
The QDO/ X-α-Gal/AbA is a laboratory reagent used for the detection and selection of recombinant proteins in bacterial and yeast expression systems. It is a combination of the following components: QDO (Quadruple Drop Out), X-α-Gal (5-bromo-4-chloro-3-indolyl-α-D-galactopyranoside), and AbA (Aureobasidin A). These components work together to enable visual identification and selection of positive clones.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using qdo x α gal aba
1
Yeast Two-Hybrid Assay for Golgin45 Binding
2
Protein-Protein Interaction Analysis of SlGRAS4
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The full‐length ORF of SlGRAS4 (Appendix S2 ) and the truncated SlGRAS4 versions were cloned into pGBDT7 vector. The recombined SlGRAS4‐pGBDT7 plasmids named SlGRAS4 (1‐667), SlGRAS4 ΔC (1‐285), SlGRAS4 ΔN1 (50‐667), SlGRAS4 ΔN2 (100‐667), SlGRAS4 ΔN3 (130‐667), SlGRAS4 ΔN4 (150‐667), SlGRAS4 ΔN5 (200‐667) and SlGRAS4 ΔN6 (286‐667) were transformed into Y2HGold strain and spread on SD/‐Trp, SD/‐Trp/X‐α‐gal and SD/‐Trp/X‐α‐gal/AbA plates for transcriptional activation test. The SlGRAS4‐pGADT7 recombined plasmid as prey and the truncated SlGRAS4 ΔN5 (200‐667) and SlGRAS4 ΔN6 (286‐667) have no transcriptional activation activity as baits were co‐transformed into Y2HGold strain and spread on SD/‐Leu/‐Trp/X‐α‐gal (DDO/X‐α‐gal), SD/‐Ade/‐His/‐Leu/‐Trp (QDO), and SD/‐Ade/‐His/‐Leu/‐Trp/X‐α‐gal/AbA (QDO/X‐α‐gal/AbA) plates for protein–protein interaction test following the manufacturer's protocol (Clontech).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!