[Generation of recombinant vaccinia virus expressing attachment glycoprotein of Nipah virus].
The mammalian condon optimized G gene was synthesized by over-lapping PCR and used to generate recombinant vaccinia virus, rWR-NiV-G. The expression of Nipah virus G protein in rWR-NiV-G infected HeLa cells was confirmed by western-blot with NiV G protein specific mouse antiserum generated by DNA immunization.The recombinant G protein showed sensitive and specific antigenic reaction to rabbit serum anti-Nipah virus in indirect florescence. Syncytium formation was induced in BHK cells by rWR-NiV-G infection following NiV F protein expressing plasmid pCAGG-NiV-F transfection. Immunization with rWR-NiV-G elicited G protein specific antibody responses in mice. The prokaryotic expressing G protein fragment showed sensitive and specific antigenic reaction to NiV G protein specific antibody from rWR-NiV-G immunized mice serum in indirect ELISA. Furthermore, the G protein specific antibodies could neutralize the infectivity of the recombinant Vesicular Stomatitis Virus pseudotype VSVAG * F/G, in which the VSV envelope protein G gene was replaced with the green fluorescent protein gene (VSVAG * G, Whitt MA) and complemented with Nipah virus F and G glycoprotein expressed in transient (VSVAG * F/G).The results here demonstrated the G protein expressed by rWR-NiV-G keeps native immunogenicity and biological activity. The recombinant virus could be promising vaccine strategy for the prevention of Nipah virus.
Wang
Xi-jun
XJ
National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China. wangxj06@yahoo.com.cn
Wang
Oing-hua
OH
Ge
Jin-ying
JY
Hu
Sen
S
Bu
Zhi-gao
ZG
D000818:Animals; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D019084:Fluorescent Antibody Technique, Indirect; D051379:Mice; D045405:Nipah Virus / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic / Q000235:genetics*; D014616:Vaccinia virus / Q000235:genetics* / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
D014759:Viral Envelope Proteins