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Volume 18 Issue 2

April 2003

Citation: YOU Yong-jin, ZHU Caizhu, GE Yan, CHEN Bo, ZHANG Qiang, RAO Zhong, XU Quan-xing, LU Yong-gan. Cloning and Expressing FM DV Non-Structural Protein 3abc Gene in E．cofi .VIROLOGICA SINICA, 2003, 18(2) : 155-158.

Cloning and Expressing FM DV Non-Structural Protein 3abc Gene in E．cofi

YOU Yong-jin ¹ ,
ZHU Caizhu ² ,
GE Yan ¹ ,
CHEN Bo ¹ ,
ZHANG Qiang ² ,
RAO Zhong ¹ ,
XU Quan-xing ¹ ,
LU Yong-gan ²

1.
Animal Genetic Engineering Research Section，Shanghai Municipal Key Laboratory ofAgricultural Genetic and Breeding， Institute ofAnima l Husbandry and Veterinary Medicine，Shanghai Academy of Agricultural Sciences，Shanghai 201106,China
2.
Lanzhou Veterinary Research Institute，China Academy of Agricultural Sciences，Lanzhou 730046,China

Available online: 15 April 2003

Abstract

Abstract：The Foot and mouth disease virus(FMDV、non．structural protein 3ABC can be used for diferentiation of infection from vaccination．The primers of 3ABC gene were designed an d synthesized， an d the 5’end an d 3’end of primers were adding the sequence of restriction endonuclease BamH I an d Hind III respectively．The 3ABC gene coding region was obtained from the FMDV genome RNA by RT．PCR．Th e amplified fragment was cloned into T．vector．The recombinan t plasmid pT．3ABC was digested with BamH I and ，ld III an d then cloned into pET32a．Th e recombinan t plasmid pET3ABC was transformed into BL21(DE3)plysS and the target protein was induced by IP1℃ ．Expression of NSP．3ABC protein was examined and identificated bV SDS．PAGE，W_estem blotting an d EU SA．Th e results showed that recombinan t plasmid pET3ABC was constructed an d the NSP．3ABC was expressed in Ecoli successfullv．A special electrophoretic ban d in SDS．PAGE (56kDa)Was noted，Westem bloting showed it Can react witIl FM DV infected an imal serum ，and ELISA result showed the expressed protein Can be used to diferentiate the infection from vaccination．
- Foot-and-mouth disease virus
- , Non-stI1lcmral protein
- , Cloning
- , Expression

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Cloning and Expressing FM DV Non-Structural Protein 3abc Gene in E．cofi

1. Animal Genetic Engineering Research Section，Shanghai Municipal Key Laboratory ofAgricultural Genetic and Breeding， Institute ofAnima l Husbandry and Veterinary Medicine，Shanghai Academy of Agricultural Sciences，Shanghai 201106,China
2. Lanzhou Veterinary Research Institute，China Academy of Agricultural Sciences，Lanzhou 730046,China

Abstract: Abstract：The Foot and mouth disease virus(FMDV、non．structural protein 3ABC can be used for diferentiation of infection from vaccination．The primers of 3ABC gene were designed an d synthesized， an d the 5’end an d 3’end of primers were adding the sequence of restriction endonuclease BamH I an d Hind III respectively．The 3ABC gene coding region was obtained from the FMDV genome RNA by RT．PCR．Th e amplified fragment was cloned into T．vector．The recombinan t plasmid pT．3ABC was digested with BamH I and ，ld III an d then cloned into pET32a．Th e recombinan t plasmid pET3ABC was transformed into BL21(DE3)plysS and the target protein was induced by IP1℃ ．Expression of NSP．3ABC protein was examined and identificated bV SDS．PAGE，W_estem blotting an d EU SA．Th e results showed that recombinan t plasmid pET3ABC was constructed an d the NSP．3ABC was expressed in Ecoli successfullv．A special electrophoretic ban d in SDS．PAGE (56kDa)Was noted，Westem bloting showed it Can react witIl FM DV infected an imal serum ，and ELISA result showed the expressed protein Can be used to diferentiate the infection from vaccination．

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