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Figure 1
Isolation of novel binding partners of HMX3 with yeast two-hybrid screening. Top: Schematic representation of bait and prey constructs used in screening. Full-length mouse HMX3 fused to the Gal4 DNA-binding domain (DBD) was used as bait. A library of stage E11 mouse cDNAs fused to the Gal4 activating domain (AD) was used as prey. 7.2 × 108 solid plate matings were performed, yielding 3200 positive colonies. Positive prey inserts were segregated and sequenced and reads were aligned to the mouse genome, identifying 851 unique genes. After removing non-coding genes and those for which only untranslated region (UTR) or intronic sequences were recovered, 539 protein-coding sequences remained. Zebrafish orthologs of the most functionally interesting genes were selected for further analysis with Co-IPs.