新城疫病毒基因Ⅲ型强毒JS/7/05/Ch的拯救

Abstract

Abstract: The genome of JS/7/05/Ch isolate shared more than 99% nucleotide identity with that of Mukteswar strain. However, the pathogenicity of JS/7/05/Ch was much stronger than that of Mukteswar strain. In order to provide a good foundation for the further related research, we built the rescue system of JS/7/05/Ch in this study. Based on the genomic sequence of JS/7/05/Ch strain, eight pairs of primers were designed to amplify the genomic fragments by RT-PCR and cloned into pCR2.1 vector, to construct the plasmid pJS/7/05/Ch which contained the full-length cDNA of NDV. Then the full-length cDNA was transferred to TVT7R(0.0) vector to construct full-length NDV infectious clone, pTVT/JS705, using specific enzymes. The infectious clone together with three helper plasmids (pCI-NP, pCI-P and pCI-L) were cotransfected into BSR-T7/5 cell, and the transfection supernatant was inoculated into SPF embryonated eggs to rescue the virus. The results of HA and RT-PCR indicated that JS/7/05/Ch strain of NDV was successfully rescued.

Key words: Newcastle disease virus; genotype Ⅲ; reverse genetics technique; rescue

CLC Number:

S852.65

SONG Qing-qing, ZHU Jie, DING Ping-yun, DUAN Zhi-qiang, TU Jie, LIU Xiao-wen, LIU Xiu-fan. Rescue of Genotype Ⅲ Velogenic Newcastle Disease Virus Strain JS/7/05/Ch[J]. , 2013, 40(10): 37-41.

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Rescue of Genotype Ⅲ Velogenic Newcastle Disease Virus Strain JS/7/05/Ch

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