title: Production and characterization of a fusion peptide derived from the rabies virus glycoprotein (RVG29).
abstract: Gene therapy targeting the brain holds great promise in curing nervous system degenerative diseases in clinical applications. With this in mind, in a previous study a 29 amino-acid peptide derived from the rabies virus glycoprotein (RVG29) with a nonamer stretch of arginine residues (RVG29-9R) at its carboxy-terminus was exploited as a ligand for brain-targeting gene delivery. Importantly, the report demonstrated that the RVG29-9R vector was able to cross the blood-brain barrier. RVG29-9R is currently synthesized by commercial companies with high associated costs. In this study, in order to reduce the costs of producing RVG29-9R, we have expressed and purified 6mg of a recombinant peptide (RVG29-9R-6His) from 0.4g of cultured Escherichia coli. We assessed the physiochemical properties of RVG29-9R-6His, its cytotoxicity, and the in vitro transfection efficiency in Neuro 2a cells (which express the acetylcholine receptor). Our results reveal that the RVG29-9R-6His peptide recognized Neuro 2a cells in a dose-dependent manner and it was also able to bind plasmid DNA and deliver it into the Neuro 2a cells effectively. Therefore, our study has demonstrated that the recombinant RVG29-9R-6His peptide retains the functions of RVG29-9R and so may provide an economically viable and alternative production method for the manufacture of RVG29-9R.
authors.affiliation: College of Animal Science and Technology, Southwest University, Chongqing 400700, China.
authors.forename: Yu-Jiao
authors.initials: YJ
authors.lastname: Yang
authors.affiliation: Military Veterinary Institute, Academy of Military Medical Sciences, Changchun 130122, Jilin Province, China; Department of Laboratory Medicine, Meizhou People's Hospital, Meizhou Hospital Affiliated to Sun Yat-sen University, Meizhou 514031, China. Electronic address: zhaopingsen@hotmail.com.
authors.forename: Ping-Sen
authors.initials: PS
authors.lastname: Zhao
authors.affiliation: Military Veterinary Institute, Academy of Military Medical Sciences, Changchun 130122, Jilin Province, China.
authors.forename: Hong-Xia
authors.initials: HX
authors.lastname: Wu
authors.affiliation: Military Veterinary Institute, Academy of Military Medical Sciences, Changchun 130122, Jilin Province, China.
authors.forename: Hua-Lei
authors.initials: HL
authors.lastname: Wang
authors.affiliation: Military Veterinary Institute, Academy of Military Medical Sciences, Changchun 130122, Jilin Province, China.
authors.forename: Li-Li
authors.initials: LL
authors.lastname: Zhao
authors.affiliation: Military Veterinary Institute, Academy of Military Medical Sciences, Changchun 130122, Jilin Province, China.
authors.forename: Xiang-Hong
authors.initials: XH
authors.lastname: Xue
authors.affiliation: Military Veterinary Institute, Academy of Military Medical Sciences, Changchun 130122, Jilin Province, China.
authors.forename: Wei-Wei
authors.initials: WW
authors.lastname: Gai
authors.affiliation: Military Veterinary Institute, Academy of Military Medical Sciences, Changchun 130122, Jilin Province, China.
authors.forename: Yu-Wei
authors.initials: YW
authors.lastname: Gao
authors.affiliation: Military Veterinary Institute, Academy of Military Medical Sciences, Changchun 130122, Jilin Province, China. Electronic address: yst610223@yahoo.com.cn.
authors.forename: Song-Tao
authors.initials: ST
authors.lastname: Yang
authors.affiliation: Military Veterinary Institute, Academy of Military Medical Sciences, Changchun 130122, Jilin Province, China. Electronic address: xiaxianzhu@gmail.com.
authors.forename: Xian-Zhu
authors.initials: XZ
authors.lastname: Xia
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D004247:DNA / Q000008:administration & dosage; D006023:Glycoproteins / Q000235:genetics*; D006801:Humans; D051379:Mice; D010446:Peptide Fragments / Q000235:genetics*; D010957:Plasmids; D055550:Protein Stability; D011820:Rabies virus / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000633:toxicity; D014759:Viral Envelope Proteins / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics*
chemical_list: D006023:Glycoproteins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C583075:rabies virus glycoprotein peptide; D004247:DNA
keywords: Brain; Gene delivery; RVG29-oligoarginine peptide; Rabies