Construction and identification of eukaryotic expression vectors of Olig1 and Id2 genes for bimolecular fluorescence complementation assay

Objective:To construct and identify pBiFC-VN173-Olig1 and pBiFC-VC155-Id2 eukaryotic expression plasmid for bimolecular fluorescence complementation (BiFC) assay.Methods:Rat olig1 gene from pGFP-N3-Olig1 eukaryotic expression plasmid was amplified by PCR.And rat Id2 gene from RNA of neonatal rat spinal cord was amplified by RT-PCR.The Olig1 gene was inserted into BiFC eukaryotic expression vector pBiFC-VN173 and Id2 gene was inserted into pBiFC-VC155,which were used to construct recombinant expression vector pBiFC-VN173-Olig1 and pBiFC-VC155-Id2,respectively.The recombinant vectors were identified by restriction enzyme digestion and DNA sequencing.Results:The restriction enzyme digestion and DNA sequencing results showed that the sequences and open read frames of the two vectors were completely concordant with experiment design.Conclusions:The pBiFC-VN173-Olig1 and pBiFC-VC155-Id2 were successfully constructed.These provide an experimental base for further research on interaction between Olig1 and Id2 in vivo.