狂犬病病毒单质粒拯救系统的建立及应用

doi:10.11843/j.issn.0366-6964.2015.11.014

Abstract

Abstract:

In order to assess the possibility of developing one-plasmid rescue systems of rabies virus(RABV)，this study was undertaken to generate a single-infectious cDNA clone by inserting Picornaviral IRES elements.Five full-length cDNA clones were constructed by inserting encephalomyocarditis virus(EMCV) IRES sequences，PV IRES sequences or TaV 2A-like sequences to upstream of the respective ORFs.BSR T7/5 cells in 6-well plates were directly transfected with 10 μg of either full-length clones or co-transfected with pTiT-N，pTiT-P or pTiT-L using CaPO₄as the control.Two recombinant RABVs were successfully rescued from these constructs，yet the rescue of SAD-NeNePeL was significantly more efficient than the rescue of SAD-NeNtPeL.Passaged SAD-NeNtPeL was genetic stable whereas passaged SAD-NeNePeL was recombinant in the RNA level.Both viruses were strongly attenuated，only growing to titers reduced about 2-log steps in comparison to the wild-type virus，SAD L16 strain.The rescue of RABVs from these single infectious cDNA clones provide a powerful tool to investigate the molecular biology，neurotropism，and pathogenicity of RABV.

CLC Number:

S852.659.5

SUN Yu-zhang，HE Biao，XU Yun-bin，CONG Yan-long，CONZELMANN Karl-Klaus. Generation of a Single Infectious RABV cDNA Clone by Inserting IRES Sequences[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(11): 2020-2024.

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Generation of a Single Infectious RABV cDNA Clone by Inserting IRES Sequences

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