Construction And Identification Of Recombinant Lentiviral Expression Vector For Human Heparanase Gene

Objective:To construct lentivirus-mediated human heparanase（HPSE）geneexpression vector and to detect its expression properties，To lay the foundationfor the test hepatoma cells influence platelet adhesion and aggregation.Methods：Human HPSE gene was synthesized and amplified by PCR,target genewas connected with Lentivirus vector Linearized by digestion,the productions ofwhich were transformed into bacterium coliDH5a cell, and positive recombinantclones were identified by PCR and analyzed directly through sequence. Then theplasmids of HPSE gene was transfected into293T cells, observed expression ofgreen fluorescence protein (GFP), Western-Blot was used to test protein express-ion of HPSE gene.At last，it was packaged into lentiviruses,and detected the titerof recombinant lentivirus Concentrate by qPCR.Results：Lentivirus expression vector of human HPSE gene was constructedsuccessfully, it was confirmed that PGC-LV-HPSE carried a right HPSE gene byfluorescence microscopy and Western-Blot, and it could be expressed in293Tcells. The titer of recombinant lentivirus was2.00E+8TU/ml tested by RT-qPCR.Conclusions：High titer recombinant lentivirus expression vector of human HPSEgene was constructed and packaged successfully.