[Construction of recombinant canine adenovirus type 2 expressing Canine coronavirus spike glycoprotein and its immunogenicity].
In order to construct a recombinant Canine adenovirus type 2 (CAV-2) expressing the spike glycoprotein of Canine coronavirus (CCV), the S1 gene fragment of CCV strain DXMV, encoding major antigenic region A, B, C and D of S protein, was amplified by RT-PCR and cloned into pVAX1 vector. The complete S1 expression cassette was subcloned into the shuttle vector pVAXE3, then further cloned into the backbone vector pPoly2-CAV2 containing complete genome of CAV-2. To gain the recombinant Canine adenovirus, the recombinant plasmid pCAV-2-CCV-S1 was linearized by Cla I/Asc I to release recombinant genome, and then transfected into MDCK cell. The recombinant virus CAV-2-S1 was gained through 4 passages in MDCK, which showed classical CPE of CAV-2. The expressed S1 protein of CCV, which was identified by RT-PCR and Western blot, can be specifically recognized by polyclonal antibody against CCV. The immunization in dogs indicated that the recombinant CAV-2 could effectively induce the specific antibodies against CCV and CAV.
Qiao
Jun
J
Institute of Military Veterinary, Academy of Military Medical Sciences of PLA, Changchun 130062, China. qj710625@yahoo.com.cn
Xia
Xian-Zhu
XZ
Yang
Song-Tao
ST
Hu
Gui-Xue
GX
Xie
Zhi-Jing
ZJ
D006516:Adenoviruses, Canine / Q000145:classification / Q000235:genetics*; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D017939:Coronavirus, Canine / Q000276:immunology*; D004285:Dogs; D007114:Immunization; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000276:immunology*; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis; D064370:Spike Glycoprotein, Coronavirus; D014614:Vaccines, Synthetic / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology*
D000914:Antibodies, Viral; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; D064370:Spike Glycoprotein, Coronavirus; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins; D014765:Viral Vaccines; C578557:spike glycoprotein, SARS-CoV; C578553:spike protein, mouse hepatitis virus