Screening proteins interacting with transcription factor ZNF580 by yeast two-hybrid system

AIM: To identify the possible proteins interacting with zinc finger transcription factor ZNF580. METHODS: The open reading frame of ZNF580 gene was amplified by PCR and ligated into plasmid pGB. The bait plasmid pGB-ZNF580 was transformed into Y190 yeast strain after sequencing. Human fetal brain cDNA library was also transformed into the bait yeast Y190 that could express the bait protein. The co-transformants were plated onto SD/-Trp/-Leu/-His selective medium. Recognition of positive colonies was performed by β-galactosidase colony-lift filter assay. Some positive plasmids were transformed into the bait yeast separately further to eliminate the false positive ones. The inserted cDNAs were extracted and analyzed for homology using the BLAST program against the GenBank/EMBL/DDBJ non-redundant sequence database. RESULTS: Fourteen proteins were identified to possibly interact with ZNF580. CONCLUSION: The possible signal pathway in which ZNF580 may be involved was initially explored.