狂犬病病毒BD-06株基质蛋白的原核表达、纯化及其多克隆抗体的制备

Abstract

Abstract: The objective of this study was to obtain polyclonal antibody against the matrix protein of rabies virus in order to provide materials for further study on the function of matrix protein. The full sequence of the matrix protein gene from the BD-06 strain rabies virus was successfully cloned and inserted into the expression vector pET28a. The results of restriction enzyme digesting and sequencing proved that the matrix protein gene was correctly inserted into pET28a. After the expression plasmid was transformed into E.coli Rosetta and induced by 0.5 mmol/L IPTG, 27 ku M protein was obtained. After the matrix protein was purified and vaccinated the rabbit,the polyclonal antibody was obtained. The results of Western blotting and IFA showed that the antibody could react with the rabies virus. The matrix protein in the BHK-21 cell affected by BD-06 strain rabies virus was positioned by using the polyclonal antibody. The results indicated that the polyclonal antibody against the matrix protein of rabies virus was successfully prepared, which laid a foundation for further study on matrix protein of rabies virus.

Key words: rabies virus; matrix protein; prokaryotic expression; polyclonal antibody

CLC Number:

ZHAO Xue-chao, ZHANG Xue-wei, ZHANG Shou-feng, LIU Ye, MI Li-juan, HU Rong-liang. Prokaryotic Expression and Purification of Matrix Protein of BD-06 Strain Rabies Virus and Preparation of its Polyclonal Antibody[J]. China Animal Husbandry & Veterinary Medicine, 2013, 40(4): 36-39.

References

1 张继希,张志远,张宜娜,等. 猪唾液酸黏附素氨基端结构域基因的克隆表达及其多克隆抗体制备[J].中国畜牧兽医,2012,39(9): 50~53.
2 徐宜为.免疫检测技术[M]. 北京:科学出版社,1991.
3 杨振慧,葛均青,龚晖,等.弹状病毒基质蛋白的研究进展[J]. 福建农业学报,2011,26(1):130~134.
4 Faber M,Pulmanausahakul R,Nagao K,et al.Identification of viral genomic elements responsible for rabies virus neuroinvasiveness[J].Proc Natl Acad Sci USA,2004,101(46):16328~16332.
5 Finke S,Conzelmann K K.Dissociation of rabies virus matrix protein functions in regulation of viral RNA synthesis and virus assembly[J]. J Virol,2003a,77(22):12074~12082.
6 Finke S,Mueller-Waldeck R,Conzelmann K K.Rabies virus matrix protein regulates the balance of virus transcription and replication[J].J Gen Virol,2003b,84(Pt6):1613~1621.
7 Finke S,Granzow H,Hurst J,et al. Intergenotypic replacement of lyssavirus matrix proteins demonstrates the role of lyssavirus M proteins in intracellular virus accumulation[J].J Virol,2010,84(4):1816~1827.
8 Golding J W.Monoclonal antibodies:Principle and practice[M].New York,Academic Press,1983.
9 Kawai A.Transcriptase activity associated with rabies viron[J].J Virol,1977,24(3):826~835.
10 Larrous F,Gholami A,Mouhamad S,et al.Two over-lapping domains of a lyssavirus matrix protein that acts on different cell death pathways[J].J Virol,2010,84(19):9897~9906.
11 Meulenberg J J, Petersen-den Besten A, De Kluyver E P, et al. Characterization of protein encoded by ORFs 2 to 7 of Lelystad virus[J].Virology,1995,206(1):155~163.
12 Pulmanausahakul R,Li J,Schnell M J,et al.The glycoprotein and the matrix protein of rabies virus affect pathogenicity by regulating viral replication and facilitating cell-to-cell spread[J].J Virol,2008,82(5):2330~2338.
13 Satoshi A,Akihiko K,Takahashi T,et al. Monoclonal antibody 3-9-16 recognizes one of the two isoforms of rabies virus matrix protein that exposes its N-terminus on the virion surface[J].Microbio Immnol,2003,47(9):639~651.
14 Wunner W H,Larson J K,Dietzschlod B,et al.The molecular biology of rabies viruses[J]. Rev Infect Dis,1988,10(suppl.4): s771~s784.

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Prokaryotic Expression and Purification of Matrix Protein of BD-06 Strain Rabies Virus and Preparation of its Polyclonal Antibody

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