title: A recombinant rabies virus carrying GFP between N and P affects viral transcription in vitro.
abstract: Several studies have demonstrated the rabies virus to be a perfect potential vaccine vector to insert foreign genes into the target genome. For this study, a green fluorescent protein (GFP) gene was cloned into the rabies virus (RABV) genome between the N and P gene. CT dinucleotide was inserted as intergenic region. The recombinant high egg passage Flury strain (HEP-Flury) of RABV, carrying GFP (rHEP-NP-GFP), was generated in BHK-21 cells using reverse genetics. According to the viral growth kinetics assay, the addition of GFP between N and P gene has little effect on the viral growth compared to the parental strain HEP-Flury. Quantitative real-time PCR (qPCR) indicated that rHEP-NP-GFP showed different viral gene transcription, especially for G gene, compared to HEP-Flury. The same is true for one other recombinant RABV carrying GFP between G and L gene in NA cells. In addition, parent HEP-Flury showed more expression of innate immune-related molecules in NA cells. Compared to HEP-Flury, Western blotting (WB) indicated that insertion of a foreign gene following N gene enhanced the expression of M and G proteins. According to the qPCR and WB, GFP expression levels of rHEP-NP-GFP were significantly higher than rHEP-GFP. This study indicates HEP-Flury as valid vector to express exogenous genes between N and P.
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Luo
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhao
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, China.
authors.forename: Qin
authors.initials: Q
authors.lastname: Tian
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, China.
authors.forename: Weiyu
authors.initials: W
authors.lastname: Mo
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, China.
authors.forename: Yifei
authors.initials: Y
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Chen
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, China. xfguo@scau.edu.cn.
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Guo
mesh_terms: D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D005822:Genetic Vectors; D016679:Genome, Viral; D049452:Green Fluorescent Proteins / Q000235:genetics*; D007113:Immunity, Innate; D018832:Molecular Chaperones; D019590:Nucleocapsid Proteins / Q000235:genetics*; D010750:Phosphoproteins / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics; D011819:Rabies Vaccines / Q000235:genetics* / Q000276:immunology; D011820:Rabies virus / Q000235:genetics* / Q000276:immunology; D014158:Transcription, Genetic; D014614:Vaccines, Synthetic / Q000235:genetics / Q000276:immunology; D019562:Viral Load; D014764:Viral Proteins / Q000235:genetics; D015678:Viral Structural Proteins / Q000235:genetics*
chemical_list: D018832:Molecular Chaperones; D019590:Nucleocapsid Proteins; C103961:P phosphoprotein, Rabies virus; D010750:Phosphoproteins; D012367:RNA, Viral; D011819:Rabies Vaccines; D014614:Vaccines, Synthetic; D014764:Viral Proteins; D015678:Viral Structural Proteins; C099601:nucleocapsid protein, Rabies virus; D049452:Green Fluorescent Proteins; C097625:L protein, Rabies virus; D012321:DNA-Directed RNA Polymerases
keywords: GFP; Gene transcription; HEP-Flury; Rabies virus; Vector