Construction and identification of recombinant adenoviruses containing fusion genes of ubiquitin and Hantaan virus Gn or Gc and S0.7
ZHANG Liang
YU Lan
YE Wei
LI Pu-yuan
ZHANG Fang-lin
CHENG Lin-feng
Abstract：Objective To construct and identify recombinant adenoviruses containing fusion genes of ubiquitin (Ub) and Hantaan virus Gn or Gc and S0.7. Methods The fragment of Ub was amplified by PCR and inserted into the pShuttle-pCAG-GnS0.7 and pShuttle-pCAG-GcS0.7 to create new transfer vectors pShuttle-pCAG-Ub-GnS0.7 and pShuttle-pCAG-Ub-GcS0.7, respectively. The pCAG-Ub-GnS0.7 and pCAG-Ub-GcS0.7 fragments were then cloned into Adeno-XTM Viral vector using PI-Sce I and I-Ceu I diges-tion. The recombinant adenovirus vectors, rAd-pCAG-Ub-GnS0.7 and rAd-pCAG-Ub-GcS0.7 linearized by PacⅠwere transfected into HEK 293 cells to package the recombinant adenoviruses. Finally, the expression of target proteins in HEK 293 cells were detected by indirect immunofluorescence assay (IFA) after being infected by recombinant adenoviruses. Results The recombinant transfer vec-tors and recombinant adenovirus vectors were confirmed by enzyme digestion, target gene PCR and sequencing. The expressions of tar-get proteins of recombinant adenoviruses were successfully detected by IFA. Conclusions The recombinant adenoviruses containing fusion genes of Ub, Hantaan virus Gn or Gc and S0.7 are successfully constructed, which can be used in further study on the immuno-logical characteristics of the Hantaan virus recombinant adenoviruses.
Keywords：Hantaan virusadenoviridaeubiquitin
Publication Date：2015-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 83-87 )