The MATCHMAKER GAL4 Two-Hybrid System 3 (Clontech) was used to examine the interaction between proteins. Appropriate amounts of bacterial solution were spread on SD/-Leu-Trp (SD/-L-T) media and incubated inverted at 30 °C for 3 days. The monoclonal antibody was picked up from SD/Leu-Trp-His-Ade (SD/-L-T-H-A) medium and resuspended in water, then diluted to a 1, 10−1, 10−2, 10−3 gradient, spotted on SD/-L-T-H-A medium, cultured at 30 °C and observed after one week52 (link).
Lu C., Du J., Chen H., Gong S., Jin Y., Meng X., Zhang T., Fu B., Molnár I., Holušová K., Said M., Xing L., Kong L., Doležel J., Li G., Wu J., Chen P., Cao A, & Zhang R. (2024). Wheat Pm55 alleles exhibit distinct interactions with an inhibitor to cause different powdery mildew resistance. Nature Communications, 15, 503.
Other organizations :
Nanjing Agricultural University, Hubei Academy of Agricultural Sciences, Institute of Plant Protection, Agricultural Institute, Centre for Agricultural Research, Czech Academy of Sciences, Institute of Experimental Botany, Shandong Academy of Agricultural Sciences
Dilution gradient of the monoclonal antibody (1, 10^-1, 10^-2, 10^-3)
dependent variables
Growth of the monoclonal antibody on SD/-L-T-H-A medium
control variables
Bacterial solution spread on SD/-L-T media
Incubation temperature of 30 °C
Incubation time of 3 days (for SD/-L-T media) and 1 week (for SD/-L-T-H-A media)
positive controls
Not explicitly mentioned
negative controls
Not explicitly mentioned
Annotations
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