The Expression, Purification Of One Kind Of Maize 14-3-3 Proteins And The Screeness Of The Protein Interacting Factors

14-3-3 proteins are popular molecular chaperon proteins in all eukaryotic cells,the recent study indicates that 14-3-3 proteins maybe interact with proteins in the path of starch metabolism in maize endosperm to regulate maize starch synthesis. In this research we designed one pair of primers according to maize GF14-12 CDs(M96856) sequence on GenBank and expand the full-length cDNA sequence that encode the GF14-12 gene in maize endosperm.The GF14-12 gene,which is one of the maize 14-3-3 protein family members was constructed into the prokaryotic expression vector pET15b::GF14-12 and the yeast two-hybrid bait vector pGBKT7::GF14-12.We induced the expression of the external GF14-12 gene in E.coli using lmM IPTG and obtained the protein through affinity column purification, providing materia for maize 14-4-3 proteins anti-body preparation for further study. Two proteins intereact with GF14-12 protein were screened out from maize endosperm using the yeast two-hybrid technology.The two proteins are Tim17 family member and Aux/IAA family member IAA6 respectively. Our study indicated that there are truly protein factors which can interact with 14-3-3 proteins in maize endosperm.