Abstract |
Hantavirus (HV) infection leads to a kind of severe systematic syndrome, hemorrhagic fever with renal syndrome ( HFRS). Heat shock proteins (HSPs) can be used as adjuvants assisting soluble antigens to produce specific targets which can be attacked by cytotoxic T lymphocytes. For further research on HFRS vaccine, this study aimed to express Hantaan virus nucleocapsid protein (HTNV NP)-HSP70 fusion protein in COS-7 cells. First, an HTNV S gene encoding NP was amplified by PCR with a mutated termination code and cloned into eukaryotic expression vector pCDNA3.1(+), into which the full-length hsp70 gene had already been inserted, to form the S-hsp70 fusion expression vector pCDNA3.1(+)/S-hsp70. Then this recombinant plasmid was transfected into COS-7 cells by liposome, and eukaryotic expression of NP-HSP70 fusion protein was detected by immunocytochemistry and western blot. The results show that the eukaryotic expression vector pCDNA3.1(+)/S-hsp70 was successfully constructed and the NP-HSP70 fusion protein was effectively expressed in COS-7 cells. This study demonstrates that the NP-HSP70 fusion protein was expressed effectively from the pCDNA3.1(+)/S-hsp70 vector in a eukaryotic system and thus provides a basis for using this plasmid as a new DNA vaccine against HV infection.
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Authors | Juan Gao, Bicheng Zhang, Shoujing Yang, Kainan Li, Hualin Xu, Yimin Xiong, Yiming Wang, Yan Wang, Weitian Xu, Yi Cheng, Guorong Zheng |
Journal | Current microbiology
(Curr Microbiol)
Vol. 58
Issue 1
Pg. 30-4
(Jan 2009)
ISSN: 1432-0991 [Electronic] United States |
PMID | 18810536
(Publication Type: Journal Article)
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Chemical References |
- Adjuvants, Immunologic
- Capsid Proteins
- HSP70 Heat-Shock Proteins
- Vaccines, DNA
- Viral Core Proteins
- nucleocapsid protein, Hantaan virus
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Topics |
- Adjuvants, Immunologic
(genetics, pharmacology)
- Animals
- COS Cells
- Capsid Proteins
(genetics, immunology)
- Chlorocebus aethiops
- Gene Expression
- HSP70 Heat-Shock Proteins
(genetics, pharmacology)
- Plasmids
- Vaccines, DNA
(genetics, immunology)
- Viral Core Proteins
(genetics, immunology)
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