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- W2388304400 abstract "Objective To construct a recombinant adenovirus encoding HS1-associated protein X-1(Hax-1)for further animal model experiments in vivo. Methods Full-length mouse-derived Hax-1 DNA was obtained by the method of RT-PCR. The Hax-1 DNA was cloned in pMD18-T, and then subcloned into pAdtrack-CMV shuttle plasmid. The product was linearized for homologous recombination with AdEasy-1 vector in BJ5183 bacteria. The positive clone was identified by restriction endonuclease digestion and confirmed by sequencing, and then the recombined adenovirus DNA was transfected into AD-293 cells for packaging and amplification of AdHax-1 virus. The virus was purified by CsCl density gradient centrifugation and the expression of Hax-1 in infected cells was monitored by EGFP fluorescence. Results Transfection of adenovirus DNA could caused cytopathic effects only in AD-293 cell but not in HeLa cell line, which proved that only the replication-defective adenovirus was produced. There was no evidence for production of wild type adenovirus. The specific expression of mouse Hax-1 was identified by PCR in AD-293 cell after infection with AdHax-1, but not in the control AD-293. Conclusion The recombinant adenovirus AdHax-1 is constructed successfully, which will be useful for investigating the role of T lymphocytes apoptosis in animal models of systemic lupus erythematosus in vivo." @default.
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- W2388304400 date "2006-01-01" @default.
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- W2388304400 title "Construction, expression, and identification of the recombinant adenovirus vector of HS1-associated protein X-1" @default.
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