Screening of proteins interacting with the cytoplasmic tail of NECL1 by yeast dual-hybrid system / 基础医学与临床

ABSTRACT

Objective To screen the proteins interact with cytoplasmic tail of NECL1.Methods The cytoplasmic tail sequence of human NECL1 gene was inserted into the "bait" vector pGBKT7 in frame.The recombinant pGBKT7-NECL1C was then transformed into yeast cells and followed by screening of human fetal brain cDNA library.Then the GST pull down assay was used to confirm the interactions between the proteins.Results Nine different sequences by sequencing the positive clones and five alternative proteins were obtained by the amino acid sequence blast.Using the GST pull down analysis,we confirmed that two of them can interact with the cytoplasmic tail of NECL1 in vitro.Conclusion The alternative proteins obtained with the yeast dual-system may interact with the cytoplasmic tail of NECL1.