The Gal-4 reporter-based ProQuest™ two-hybrid system (Invitrogen, Darmstadt, Germany) was employed to identify the interactions between MAPKs and MAPKKs. Coding sequences (CDSs) of all the MAPK members of Musa were cloned into the prey vector (pDEST32), and CDSs of MAPKKs were introduced into the bait vector (pDEST22). All the combinations of each bait and prey plasmid were PEG-transformed into the yeast strain MaV203 following the protocol of the ProQuest™ two-hybrid system. Positive transformants were first chosen in the synthetic dropout (SD) medium without leucine and tryptophan (SD/−Leu/−Trp), and the culture was transferred to the selection medium without leucine, tryptophan, histidine, and adenine. 3-Amino-1,2,4-triazole (3-AT) was supplemented to the selection plates to inhibit the auto-activation of the prey vectors.

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.