猪传染性胃肠炎病毒M、sM基因重组杆状病毒的构建及病毒样颗粒的组装

doi:10.11843/j.issn.0366-6964.2013.09.014

Abstract

Abstract:

Recombinant baculovirus containing M and sM gene of transmissible gastroenteritis virus (TGEV) were constructed using Bac-To-Bac Baculovirus expression system, and recombinant M, sM proteins were expressed by infecting the sf9 cell with recombinant baculovirus. Firstly, M gene and sM gene of TGEV were amplified, then were cloned and inserted into donor plasmid pFastBacTMDual to obtain recombinant donor plamid, pFastBac^TMDual-M and pFastBac^TMDual-M-sM. Plasmid pFastBac^TMDual-M and pFastBacTMDual-M-sM were transformed into E. coli DH10Bac respectively. Recombinant baculovirus rBac-M and rBac-M-sM were obtained by transfection of the sf9 cells with Bacmid-M and Bacmid-M-sM. Western blot and indirect immunofluorescence assay (IFA) were conducted, and the results showed that recombinant M protein and sM protein was expressed successfully, two proteins had a positive reaction with antiserum. In vitro experiments of virus-like particles (VLP) assembly of TGEV was performed by infection of insect cell with rBac-M and rBac-M-sM. It is showed that the expressed M proteins alone and co-expression of M and sM proteins in insect cell Sf9 infected by recombinant baculovirus could form VLPs morphologically similar to virion of TGEV. Diameter of VLP is variable from 61 nm to 101 nm. This study provides the foundation for researching the VLP assembly of TGEV.

CLC Number:

S852.659.6

LENG Yong, SONG Zhen-hui, QING Jia-chao, ZHAI Shao-hua, MAIMAITI-Aizizi. Expression of M Gene and sM Gene of Porcine Transmissible Gastroenteritis Virus by Baculovirus Expression System and VLPs Assmbly of TGEV[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(9): 1432-1437.

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Expression of M Gene and sM Gene of Porcine Transmissible Gastroenteritis Virus by Baculovirus Expression System and VLPs Assmbly of TGEV

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