Abstract:

[Abstract] Objective   To overcome the fact that SRV- NM virus can only multiple and amplify through partially pu?
rified jaagsiekte retrovirus inoculated intratracheally in sheep but it cannot be augmented usingin vitro cell culture, we con?structed JSRV-NM pseudovirions based on high efficiency packing system of lentivirus.Methods   Lentivirus of three highefficiency packing plasmids system pMD.G, pCMV-HIV8.2and pHIV-eGFP was developed, and JSRV-NM-env coatedplasmid pCMVJSRV-NM was used to substitute VSV-G virus coated plasmid pMD.G then co-transfected into293T cells toreplicate, package and produce restructured JSRV-NM pseudovirions.Gene expressionofpseudovirion was determinedthrough WPRE using real time PCR; Virus infectivity was detected through inoculating JSRV- NM pseudovirions into 24pore plates.Results   We construct JSRV-NM pseudovirions successfully based on the lentivirus system. JSRV-NM pseudo?virions can also be concentrated to higher titer (108TU/mL detected by real time PCR by ultracentrifugation without signifi?ant loss of activity. JSRV-NM and VSV-G pseudovirions infected on Hela cells (both MOI =3) respectively and no obvi?ous difference were shown on their infection efficiency detected by real time PCR.Conclusion   Based on lentivirus system,JSRV-NM pseudovirions can be multipled and amplified in293T cell culturein vitro. JSRV-NM pseudovirions is stablewithout loss its infection activity and the requirements of biological laboratory safety II was also met. JSRV-NM pseudoviri?ons will provide a useful tool for further study of JSRV-NM–env infection across species or its induction of lung adenocarci?noma.

Key words: JSRV-NM viruses, pseudovirions, lentiviral vector, env gene, virus packageing