Using rabies virus vaccine strain SRV9 as viral vector to express exogenous gene.
Rabies virus (RABV) can cause a fatal neurological disease in human and animals, and vaccines were generally applied for the immunoprophylaxis of rabies. Here, a recombinant viral vector carrying the exogenous gene expression component between phosphoprotein (P) and matrix protein (M) genes of RABV was constructed based on the vaccine strain SRV9 used in China. To develop a reverse genetic system, the full-length cDNA plasmids of SRV9 were constructed using the eukaryotic expression vector pCI or pcDNA3.1(+). However, recovery efficiency based on the pcDNA3.1 vector was significantly higher than that of the pCI vector. The exogenous gene expression component PE-PS-BsiWI-PmeI or PS-BsiWI-PmeI-PE was introduced in different locations between the P and M genes of SRV9. When the enhanced green fluorescent protein (eGFP) was used as a reporter gene, both locations could rescue recombinant RABV (rRABV) expressing eGFP with high efficiency. Characterization of rRABV expressing eGFP in vitro revealed that its growth was similar to that of the parental virus. Animal experiments showed that rRABV expressing eGFP could replicate and express eGFP in the brains of suckling mice. Furthermore, rRABV of SRV9 was nonpathogenic for 3-week-old mice and could be cleared from the central nervous system at 5 days post-inoculation. Our results showed that the recombinant SRV9 virus could be used as a useful viral vector for exogenous gene expression.
Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, 130122, China.
Hualei
H
Wang
Hongli
H
Jin
Na
N
Feng
Xuexing
X
Zheng
Ling
L
Li
Yinglin
Y
Qi
Meng
M
Liang
Yongkun
Y
Zhao
Tiecheng
T
Wang
Yuwei
Y
Gao
Changchun
C
Tu
Ningyi
N
Jin
Songtao
S
Yang
Xianzhu
X
Xia
D000818:Animals; D005260:Female; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011819:Rabies Vaccines / Q000235:genetics* / Q000378:metabolism; D011820:Rabies virus / Q000235:genetics* / Q000378:metabolism; D059386:Reverse Genetics
D011819:Rabies Vaccines; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins