Screening Of Proteins Interacting With The SPAG6by Yeast Two Hybrid System

Objective: To explore the proteins which can interact with SPAG6, construct therecombinant plasmid of SPAG6/pGBKT7and pick the proteins by yeast two hybrid.Methods:1.Looking for the CDS of SPAG6of mice in PubMed;2.Using the testiscDNA of mice as a model, amplify SPAG6sequence by PCR;3.TA cloning;4.Construct the recombinant plasmid of SPAG6/PGBKT7;5.Transform SPAG6/pGBKT7into AH109competent cell, making yeast two hybrid with cDNA library of mice;6.Extracting plasmids of yeasts;7.Transform yeast plasmids into DH5a;8.Sequenceanalysis of these plasmids.Results: The results of restriction enzyme digestion and sequencing indicate therecombinant plasmid of SPAG6/pGBKT7construct successfully. In yeast two hybrid,initial screen some proteins which may have interaction with SPAG6.Conclusion: There are many proteins may interacting with SPAG6. This provides abasis for how to research the proteins interact with SPAG6.