Establishment Of Transgenic Fly That Expresses Recombinant Protein In Drosophila: For Analysis Of Biochemical Functions Of The Protein Complex

4 Objective To analyze biochemical functions of PCL protein in different embryogenetic development, we express PCL recombination protein complex in Drosophila, then detect and purify the protein complexes from in vivo materials.Methods PCL gene was amplified by PCR and directly inserted into pCaS83 HHF_PresCT vector at NotI/StuI sites. The expression vector of pCaS83 PCL_e was injected into Drosophila embryos to establish transgenic flies expressing epitope-tagged recombinant PCL protein. Judging from genetic and immuno-fluorescence analyses, immuno-affinity purification of the protein complex by aid of anti-FLAG antibody revealed that our methods would be applicable to some biochemical analyses.Results We established the transgenic fly that expresses recombinant protein in Drosophila. Different transgeneic fly line has different PCL protein expression level, but expressed recombinant PCL protein complex apparently behaved as endogenous PCL proteins, further more, it could partially compensated the function of detected PCL protein. For recombination PCL protein has C-terminal 8xHis-HA-FLAG epitopes, we can use anti-FLAG antibody to purify PCL complex from embryogenetic development.Conclusion We have successfully established the transgeneic fly line to express recombination PCL protein, and which, without constraint, prompted us to analyze further detail of biochemical functions of protein complex of interest.