Screening And Identification Of Interaction Proteins With LOC401296

Objectives By yeast two hybrid screen out the interacting protein of LOC401296 molecules, and through which to study the function of LOC401296 molecules.Methods 1 Screening the interaction protein of LOC401296 by yeast two hybrid method. First build the p DBLeu-loc fusion expression vector, transferred it into yeast strain Ma V203. Detected the self activation and toxic of it. Then screen adult liver c DNA library. Finally, rotary verify the positive clone, to exclude false positive clones. Analyze the screened positive clone by bioinformatics. 2 By using the method of immunofluorescence combined with confocal laser technology to observe the co LOC alization of interaction between LOC401296 and protein in HEK293 cells.Results 1 By yeast two hybrid screening out LOC401296 interacting proteins. Successfully constructed the p DBLeu-loc fusion expression vector. Rotary verified the positive clones screened. The positive genes obtained by bioinformatics analysis, geting a total of 8 positive clones. 2 Successfully constructed three fusion expression vector, p EGFP-GRN, p EGFP-PLSCR1, p EGFP-N4BP2L2. By using the method of immunofluorescence combined with confocal laser technology, observation of LOC401296 respectively with GRN, PLSCR1, N4BP2L2 in HEK293 cells co LOCalized. There were found co LOCalization of LOC401296 with 3 interacting protein.Conclusions 1 Screening the interaction protein of LOC401296 by yeast two hybrid technology, 8 positive clones were screened: GRN, PLSCR1, N4BP2L2, FN1, NOTCH3, LTBP3, ADAMTS-L4, EGFL7. These genes have different functions, which suggests that LOC401296 is involved in the process of the complexity of biological diversity and its mechanism of action. 2 Through the method of immunofluorescence combined with confocal laser technology for LOC401296 and GRN, PLSCR1, N4BP2L2 were studied in cellular LOCalization, observed the expression of LOC401296 and GRN, PLSCR1, N4BP2L2 in HEK293 cells and their good co LOCalization. These provides clues for further study of LOC401296 function.