Screening For Proteins That Can Interact With PhrIP1 By Yeast Two-hybrid System

PhrIP1(phragmoplastin-interacting proteint l),a protein of 608 amino acids,appears to play an important role in the formation of cell plate during cytokinesis in plants. The study on the function and mechanism of PhrIP1 is important for clarifying how the cell plate and cell wall form in plants.In this study,full length of phrIP1 is amplified by PCR and ligated into pKS plasmid,then the bait plasmid,pEG202-phrIP1,is constructed.The inseret gene are sure to be translated into the right fusion protein through its sequence.In the yeast two-hybrid system,the bait plasmid (pEG202-phrIP1) and a reporter plasmid (pSH18-34) are introduced into the yeast (EGY48) by co-transformation.Then cDNA library (which is in pJG4-5)is screened and two genes are obtained.The two insert gene fragments are sequenced.One of them is plastocyanin,the other is putative photosystem I reaction center subunit II precursor,both of them are the necessary components of photosynthetic chain. The more experiments should be taken to identify if they have ture interaction with PhrIP1.