Objective   To construct the endothelial cell model with overexpressed human protein kinase C β

2

(PKCβ

2

) after high glucose inducement in order to study the function of human PKCβ

2

.    Methods   The PRKCB1 gene was amplified from pMD18-T-PRKCB1 plasmid and then directly cloned into shuttle plasmid pDC315 to construct shuttle plasmid. Then the recombinant shuttle plasmid and adenovirus genomic plasmid pBHGlox△E1

3Cre were co-transfected into 293

cells to construct recombinant adenovirus Ad-PRKCB1. The virus titer was calculated by TCID50. The Ad-PRKCB1 was verified by immunocytochemistry and RT-PCR. Ad-PRKCB1 was transfected into human umbilical vein endothelial cells (HUVECs) followed by the treatment of high glucose (25 mmol/L) for 96 h

and then the cells were observed by laser scanning confocal microscopy (LSCM).    Results   Restrictive endonuclease digestion and PCR result showed that the target gene was correctly cloned into shuttle plasmid. Cytopathic effect (CPE) was observed under inverted microscope through homologous recombination. The virus titer was 7.9×10

9

 IU/ml. Immunocytochemical staining and RT-PCR indicated the expression of human PKCβ

2

in the transfected HUVECs. The translocation was observed under LSCM.    Conclusion   A recombinant adenovirus vector with human PKCβ

2

and the endothelial cell model with human PKCβ

2

overexpression by high glucose are constructed successfully.