Screening of proteins interacting with LRP16 by yeast two-hybrid

Objective:To screen LRP16 interaction proteins from cDNA library of MCF-7 human breast cancer cell line by yeast two-hybrid.Methods: The library was constructed and screened by cotransformation of the double strand cDNA and the linearized pGADT7-Rec with the bait into the AH109 yeast strain.The inserted fragments of identified positive clones were amplified by PCR and sequenced.And library plasmids of positive clones were obtained from competent bacterium coli Top10 transformated with yeast plasmid.Following,the interactions of LRP16 and positive plasmid were tested in yeast cells again.Results:There were 8 different candidate proteins obtaind by Blast analysis in NCBI.Four putative positive library plasmids were further tested interaction in yeast by transformation and three could group yeast clones.Conclusions:We obtain a novel class of LRP16 interacting proteins by yeast two-hybrid system.These results provide essential clues for further investigation of the functional regulation of LRP16 and identification.