The construction of recombinant lentiviral-vector with human interleukin-10 gene
- VernacularTitle:人白细胞介素10基因慢病毒载体的构建及重组
- Author:
Zhenghua HE
;
Nianyue BAI
;
Qulian GUO
- Publication Type:Journal Article
- Keywords:
Lentiviral-vector;
Human IL-10;
Therapy,Gene
- From:
Chinese Journal of General Surgery
2001;0(08):-
- CountryChina
- Language:Chinese
-
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?1010)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.
