Construction And Expression Of Recombinant Plasmids

Objective To respectively construct eukaryotic red fluorescence protein expressing recombinant plasmids pERFP-CI-p100,pERFP-CI-SN and pERFP-CI-TSN,which contain human p100 full length cDNA,SN fragment cDNA,TSN fragment cDNA.Methods The p100 full length cDNA,SN fragment cDNA and TSN fragment cDNA were amplified by PCR.The three kinds of foreign fragments were cloned to the eukaryotic expression vector pERFP-CI and constructed the homologous constitutive plasmids.And then They were inserted into pEGFP-CI fluorescent expressing vector.We detected the aim lanes,p100 full length,SN fragment and TSN fragment by double digestion.Transfect these recombinant plasmids into HeLa cell line and observe the expression of fluorescent proteins.Resultsâ‘ we can get the p100 full length and its fragments through the PCRâ‘¡The fragment,p100 full length,was detected in the products of the restriction enzyme digestion.â‘¢Through the blue-white spot screening,the fragments,SN and TSN, were also detected in the products of the restriction enzyme digestion.â‘£The red fluorescent proteins could be observed by fluorescence microscope after the transfection.Conclusion The fluorescent expressing recombinant plasmids,which expressed p100-RFP,SN-RFP,TSN-RFP fusion proteins,were constructed successfully.