Abstract:Objective:To construct contains human interleukin-10 gene recombinant lentiviral-vector (LV-IL-10) and to form a basis to further explore the therapy of chronic pain.
Methods :hIL-10 gene fragment was isolated and amplified from pCYIL-10 plasmid by PCR, and was cloned into pWPXL-GFP. The inserted hIL-10 fragment was verified by Pme I digestion and DNA sequencing. The recombinant plasmid pWPXL-IL-10-GFP, envelope plasmid pMD2.G and packaging plasmid psPAX2 were cotransfected into 293T cells, to pack out lentivirus particle that has the ability of duplicated-deficiency, then virus titer determination was undertaken.
Results:The 530bp IL-10 gene fragment was amplified from pCYIL-10 plasmid by PCR, and was recombinated into pWPXL-GFP plasmid. DNA sequencing confirmed that the cloned gene segment was 100% homologous to the published hIL-10 sequence in genebank. High titer（2×1010）and highly purified lentiviral particles was obtained.
Conclusions:The lentivirus vector LV-hIL-10 was constructed successfully, which form a basis of research of chronic pain therapy.