[1]高慎阳,邹运明,李一经*.猪流行性腹泻病毒M基因及其片段原核表达效果分析[J].中国预防兽医学报,2007,(01):27-31.
 GAO Shen-yang,ZOU Yun-ming,LI Yi-jing*.Prokaryotic expression analysis of recombinant M gene and its truncated M’ fragment of porcine epidemic diarrhea virus[J].Chinese journal of preventive veterinary medicine,2007,(01):27-31.
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猪流行性腹泻病毒M基因及其片段原核表达效果分析
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2007年01期
页码:
27-31
栏目:
病原生物学
出版日期:
2007-01-01

文章信息/Info

Title:
Prokaryotic expression analysis of recombinant M gene and its truncated M’ fragment of porcine epidemic diarrhea virus
文章编号:
1008-0589(2007)01-0027-05
作者:
高慎阳邹运明李一经*
东北农业大学 动物医学院,黑龙江 哈尔滨 150030
Author(s):
GAO Shen-yang ZOU Yun-ming LI Yi-jing*
Department of Veterinary, Northeast Agricultural University, Harbin 150030, China
关键词:
猪流行性腹泻病毒重组M蛋白原核表达
Keywords:
porcine epidemic diarrhea virus recombinant M protein prokaryotic expression.
分类号:
S852.65+9.6
文献标志码:
A
摘要:
将猪流行性腹泻病毒的M基因及编码M蛋白膜外区M基因片段(M’)分别亚克隆到原核表达载体pJLA605和pGEX-6p-1中,构建了重组质粒pJLA605-M、pGEX-6p-M-GST、pGEX-6p-M和pGEX-6p-M’,转化大肠杆菌后,探索了M蛋白在N端和C端不带有谷胱苷肽巯基转移酶(GST)、N端和C端均带有GST、只在N端带有GST以及只含有M蛋白的膜外区M’蛋白的表达情况。SDS-PAGE分析结果表明,N端和C端带有GST的pGEX-6p-M-GST和只含有膜外区M’基因的重组质粒pGEX-6p-M’ 获得了高效表达,经Bandscan5.0软件分析,其表达产物分别以融合蛋白GST-M和GST-M’ 形式存在,表达量分别为20 %和45 %。在对重组质粒pJLA605-M转化的大肠杆菌诱导表达过程中发现菌体生长的抑制现象,说明M蛋白对宿主菌有一定的毒性,进一步提取包涵体进行SDS-PAGE发现M蛋白以包涵体的形式得以表达。猪流行性腹泻病毒M蛋白的毒性作用可能与M蛋白具有的出芽功能有关,因而表现出对大肠杆菌的细胞壁的破坏作用。
Abstract:
The whole M gene of porcine epidemic diarrhea virus (PEDV) and its truncated fragment M’ gene which encodes the exterior of the viral envelope, were subcloned into pJLA605 and pGEX-6p-1, respectively. The recombinant plasmids pJLA605-M, pGEX-6p-M-GST, pGEX-6p-M and pGEX-6p-M’ were constructed and transformed into E.coli for expression. SDS-PAGE analysis showed recombinant M protein and truncated M’ protein were highly expressed by pGEX-6p-M-GST and pGEX-6p-M’, and the expression products as fusion protein GST-M and GST-M’ account reached 20 % and 45 % in the whole bacteria protein, respectively. With the analysis of software Bandscan5.0 and SDS-PAGE, the extract of inclusion body (IB) confirmed M proteins were expressed in the form of IBs. In addition, we found the phenomenon of growth inhibition of E.coli anchored plasmid pJLA605-M after induced,which suggest that M proteins were toxic for host bacteria, and the toxic effect may be contributed to the sprout function of M proteins and causes damage to the cell wall of E.coli.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期:2005-12-13
基金项目:黑龙江省“十五”科技攻关计划项目(GC01B510)
作者简介:高慎阳(1979~),男,黑龙江省五大连池市人,硕士研究生,主要从事预防兽医学研究.
*通讯作者:E-mail:yijingli@yahoo.com
更新日期/Last Update: 2011-01-31