Screening Interactive Proteins of CLN8P by Yeast Two Hybrid Techniques

Objective To screen and identify the interactive proteins of CLN8P and provide clues for the studies on CLN8 function and pathogenesis of NCLs. Methods The Human Fetal Brain cDNA library was screened with with pLexA-CLN8 as bait plasmid by yeast-two hybrid system, and Leu+ and LacZ+ positive yeast clones were obtained, which were cotransformed to the EGY48 (p8op-LacZ) along with pLexA-CLN8 plasmid one to one by yeast simultaneous cotransformation so as to identify the protein interactions again. The inserted fragments of identified positive clones were sequenced and analyzed with Blast in NCBI. Results There are 60 Leu+ and LacZ+ positive yeast clones which are obtained through screening the fetal brain library with pLexA-CLN8 as bait plasmid. There are 22 positive clones identified one to one by yeast simultaneous co-transformation. 5 different candidate gene sequences are obtained by Blast analysis in NCBI. Conclusion 5 gene sequences were obtained, these coding proteins can interact with CLN8P,and some of them may be involved with pathogenesis of NCLs.