首页|期刊导航|中华医学杂志（英文版）|Recombinant adenovirus with human indoleamine-2,3-dioxygenase and hepatitis B virus preS was constructed and expressed in HepG2 cells
中华医学杂志（英文版）2011，Vol.125Issue(19)：3159-3163,5.DOI:10.3760/cma.j.issn.0366-6999.2011.19.036
Recombinant adenovirus with human indoleamine-2,3-dioxygenase and hepatitis B virus preS was constructed and expressed in HepG2 cells
Recombinant adenovirus with human indoleamine-2,3-dioxygenase and hepatitis B virus preS was constructed and expressed in HepG2 cells
摘要
Abstract
Background Indoleamine-2,3-dioxygenase (IDO) is proven to suppress hepatitis B virus (HBV) specific immune response and depletion of IDO may be a useful approach for HBV therapy. To test this concept, we constructed recombinant adenovirus with human IDO and HBV preS, which would form the basis for future in vivo experiments.Methods The fragment of human IDO and HBV preS cDNA were subcloned into multiple cloning sites in an adenoviral vector system containing two cytomegalovirus (CMV) promoters. Recombination was conducted in the Escherichia coli BJ5183. The recombinant adenovirus containing hlDO gene and HBVpreS gene was packaged and amplified in 293 cells.Integration was confirmed by polymerase chain reaction as well as the quantification of viral titers. HepG2 cells were infected with the recombinant adenovirus and mRNA and protein specific for hlDO and HBVpreS was detected by RT-PCR and Western blotting respectively.Results The recombinant adenovirus was produced successfully. Its titer was 2.5x109 efu/ml. IDO and HBVpreS mRNA as well as the encoded proteins could be found in transfected HepG2 cells, but not in control HepG2 cells.Conclusion The transfer of hlDO-HBVpreS with double-promoter adenoviral vector was efficient. The recombinant adenovirus with hlDO and HBVpreS would provide the experimental basis for future studies.关键词
indoleamine-2,3-dioxygenase/recombined adenovirus/hepatitis B virus/HepG2 cellKey words
indoleamine-2,3-dioxygenase/recombined adenovirus/hepatitis B virus/HepG2 cell引用本文复制引用
CHEN Yong-bing,SHI Xian-jie,LU Gang,NIE Hong-feng,SHEN Xiao-qing,YU Cong-hui,GONG Jian-ping..Recombinant adenovirus with human indoleamine-2,3-dioxygenase and hepatitis B virus preS was constructed and expressed in HepG2 cells[J].中华医学杂志（英文版）,2011,125(19):3159-3163,5.基金项目
This work was supported by a grant from the China Postdoctoral Science Foundation (No.20060390678). （No.20060390678）
