Objective     To construct Neuropilins-2 eukaryotic expression vector for RNA interference.      Methods     Recombinant targeting on gene NRP2 was designed and established with plasmid pGenSil-1 based on NRP2 cDNA equences of Genomes. Two pairs of oligonucleotides were synthesized according to the Tuschl and inserted into plasmid pGenSil-l to generate siRNA eukaryotic expression vector

DH5α strains were transformed

plasmid were extracted

and recombinant vectors were identified by the restriction map and the sequence analysis. The recombinant plasmid (pGenSil-NRP2) was transfected into the cultured LOVO cells. At 48 h after transfection

the whole cell protein was extracted

and the protein level was detected by Western blotting with mouse-anti-human NRP2 monoclonal antibody.      Results     Recombinant plasmids were completely coincided with the designs by the restriction map and the sequence analysis. pGenSil-NRP2 expression vector into LOVO cells down-regulated the protein level of NRP2 at 48 h after transfection. The recombinant eukaryotic expression vector were constructed successfully.      Conclusion     siRNA recombinant can be constructed successfully by RNAi technique for inhibiting NRP2 expression.