全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

河南猪流感病毒H3N2分离株NP基因表达及抗原性分析

DOI: 10.11847/zgggws2015-31-09-19, PP. 1173-1175

Keywords: 猪流感病毒,NP基因,表达,抗原性

Full-Text   Cite this paper   Add to My Lib

Abstract:

?目的表达猪流感病毒(swineinfluenzavirus,SIV)H3N2分离株NP基因,获得具有良好抗原性的NP蛋白。方法PCR扩增猪流感病毒A/swine/Henan/2/2008(H3N2)NP基因,经EcoRI和XhoI双酶切后插入pET32a(+)载体,并转化大肠杆菌Rosset(DE3)菌株;PCR、EcoRI和XhoI双酶切及DNA测序鉴定重组质粒pET32a-NP;IPTG诱导,表达、纯化NP并免疫家兔制备免疫血清,SDS-PAGE、WB测其反应原性;同时构建pcDNA3.1-NP并转染293T细胞,免疫荧光检测其免疫源性。结果成功构建重组载体pET32a-NP和pcDNA3.1-NP,SDS-PAGE和WB显示融合蛋白约80kD,能与感染SIV的猪血清特异性反应,并且纯化蛋白制备的多克隆血清能识别293T细胞中表达的NP蛋白。结论正确表达SIVH3N2NP蛋白,并有良好的抗原性,为研究猪流感病毒NP诊断抗原和亚单位疫苗奠定基础。

References

[1]  赵朴,郑玉姝,李海燕.猪流感—一个不容忽视的人兽共患病[J].中国兽医杂志,2008,44(9):54-56.
[2]  Smith GJ,Vijaykrishna D,Bahl J,et al.Origins and evolutionary genomics of the 2009 swine-origin H1N1 influenza A epidemic[J].Nature,2009,459(7250):1122-1125.
[3]  Heiny AT,Miotto O,Srinivasan KN,et al.Evolutionarily conserved protein sequences of influenza a viruses,avian and human,as vaccine targets[J].PLoS ONE,2007,2(11):e1190.
[4]  赵朴,郑玉姝,贾贝贝,等.猪流感病毒HA基因的克隆及杆状病毒转移载体的构建[J].生物技术通报,2009,(9):97-100.
[5]  赵朴,郑玉姝,贾贝贝,等.H3N2 SIV 河南株NS1重组蛋白的纯化及Dot-ELISA的初步建立[J].西南师范大学学报:自然科学版,2011,36(3):190-193.
[6]  Ito T,Couceiro J,Kelm S,et al.Molecular basis for the generation in pigs of influenza A virus with pandemic potential[J].J Virol,1998,72:7367-7373.
[7]  Brown IH,Harris PA,McCauley JW,et al.Multiple genetic reassortment of avian and human influenza A viruses in European pigs,resulting in the emergence of an H1N2 virus of novel genotype[J].J Gen Virol,1998,79(Pt 12):2947-2955.
[8]  龚震宇,陈恩富,吕华坤,等.人感染甲型H7N9禽流感疫情及动物源性疾病防控分析[J].中国公共卫,2014,30(1):35-37.
[9]  Zhu X,Yu W,McBride R,et al.Hemagglutinin homologue from H17N10 bat influenza virus exhibits divergent receptor-binding and pH-dependent fusion activities[J].Proc Natl Acad Sci USA,2013,110(4):1458-1463.
[10]  Carragher DM,Kaminski DA,Moquin A,et al.A novel role for non-neutralizing antibodies against nucleoprotein in facilitating resistance to influenza virus[J].J Immunol,2008,181(6):4168-4176.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133