Amplicon sequencing
A cDNA library of N. crassa for Y2H was constructed by Gateway recombination technology. Using the MSP-8 CDS constructed on the pGBKT7 vector as bait (no self-activation), the cDNA library of N. crassa was screened. After screening on SD/-Trp/-Leu (SD-2) or the selective medium SD/-Trp/-Leu/-His/-Ade (SD-4), DNA sequencing, and BLAST comparison analysis of positive clones, the proteins that interact with pGBKT7-MSP-8 were identified. To thoroughly screen the interacting proteins, all positive clones of the yeast screening library were scraped with 2YPDA liquid, centrifuged and collected for colony PCR, and the PCR products were subjected to NGS sequencing.