Sequencing and rescuing a highly virulent classical swine fever virus:   Chinese strain cF114 from a full-length cDNA clone

doi:10.1360/03wc0006

CORC > 北京大学 > 生命科学学院

	Sequencing and rescuing a highly virulent classical swine fever virus: Chinese strain cF114 from a full-length cDNA clone
	Nie, YC ; Chen, JG ; Ding, MX
刊名	科学通报英文版
	2003
关键词	classical swine fever virus full-length cDNA cloning infectious RNA transfection runoff transcription INFECTIOUS RNA MAJOR DETERMINANT RECOVERY PESTIVIRUS PROTEIN GENOME
DOI	10.1360/03wc0006
英文摘要	The complete nucleotide sequence of classical swine fever virus (CSFV) strain cF114 (F114 strain propagated on PK-15 cells) was cloned by RT-PCR. The analyses of nucleotide and amino acids identity between cF114 and F114, Brescia, Alfort or C strain were 99.41%, 96.80%, 86.03%, 95.70% and 99.28%, 98.54%, 93.33%, 97.41% respectively. The cDNA fragments with correct sequence were ligated into a full-length cDNA and inserted into pMC18 plasmid (pMC12297). A full-length infectious viral RNA was synthesized by runoff transcription and transfected to PK15 cells. Viruses were recovered from transfected cells which wese titrated on PK-15 cells by endpoint dilution and indirect immunofluorescence with a CSFV-specific monoclonal antibody. The antigenicity and replication kinetics of the plasmid-derived virus (vM12297) were similar to the parental virus in vitro. The E01 or E2 gene was replaced with the genes from strain C and the pM/CE01 and pM/CE2 with chimeric full-length cDNA of cF114 were generated. The infectious viruses were obtained from pM/CE01 and pM/CF2. Both of the chimeric viruses can infect PK-15, SK-6 and primary testicle cell of swine. The chimeric viruses can grow to a titer of 8X10(5) F-PFU/mL. These results are very important for understanding the genes related to the CSFV propagation and pathogenesis.; http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000183662600015&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=8e1609b174ce4e31116a60747a720701 ; Multidisciplinary Sciences; SCI(E); 1; ARTICLE; 11; 1124-1128; 48
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/255873]
专题	生命科学学院
推荐引用方式 GB/T 7714	Nie, YC,Chen, JG,Ding, MX. Sequencing and rescuing a highly virulent classical swine fever virus: Chinese strain cF114 from a full-length cDNA clone[J]. 科学通报英文版,2003.
APA	Nie, YC,Chen, JG,&Ding, MX.(2003).Sequencing and rescuing a highly virulent classical swine fever virus: Chinese strain cF114 from a full-length cDNA clone.科学通报英文版.
MLA	Nie, YC,et al."Sequencing and rescuing a highly virulent classical swine fever virus: Chinese strain cF114 from a full-length cDNA clone".科学通报英文版 (2003).