| 传染性胰腺坏死病毒VP3基因的分段表达及其抗原表位区域分析 |
| |
| 引用本文： | 刘巍巍,赵丽丽,赵永欣,王健楠,李一经,葛俊伟,乔薪瑗,刘敏.传染性胰腺坏死病毒VP3基因的分段表达及其抗原表位区域分析[J].淡水渔业,2011(4):61-65. |
| |
| 作者姓名： | 刘巍巍 赵丽丽 赵永欣 王健楠 李一经 葛俊伟 乔薪瑗 刘敏 |
| |
| 作者单位： | 东北农业大学动物科学与技术学院;东北农业大学动物医学学院 |
| |
| 基金项目： | 黑龙江省自然科学基金(C200837);黑龙江省教育厅科技项目(11541019) |
| |
| 摘 要： | 采用PCR方法克隆了传染性胰腺坏死病毒(IPNV) VP3四段相互重叠的基因片段L1、L2、L3和L4,将PCR产物分别连接到原核表达载体pGEX-6P1和pET32a上,经酶切、PCR、测序鉴定,获得重组质粒pGEX-6P1 -VP3(L1)、pET32a-VP3( L2)、pGEX-6P1-VP3( L3)和pGE...
|
| 关 键 词： | 传染性胰腺坏死病毒 VP3基因 原核表达 抗原表位 |
Expression of overlapping fragments of IPNV VP3 gene and analysis of antigenic epitope domains |
| |
| LIU Wei-wei,ZHAO Li-li,ZHAO Yong-xin,WANG Jian-nan,LI Yi-jing,GE Jun-wei,QIAO Xin-yuan,LIU Min.Expression of overlapping fragments of IPNV VP3 gene and analysis of antigenic epitope domains[J].Freshwater Fisheries,2011(4):61-65. |
| |
| Authors: | LIU Wei-wei ZHAO Li-li ZHAO Yong-xin WANG Jian-nan LI Yi-jing GE Jun-wei QIAO Xin-yuan LIU Min |
| |
| Affiliation: | 1(1.College of Animal Science and Technology,Northeast Agricultural University,Harbin 150030;2.College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030) |
| |
| Abstract: | Four overlapping fragments L1,L2,L3 and L4 of VP3 gene of infectious pancreatic necrosis virus(IPNV) were amplified by PCR.PCR products were cloned into the expression vector pGEX-6P1 and pET32a.The recombinant plasmids pGEX-6P1-VP3(L1),pET32a-VP3(L2),pGEX-6P1-VP3(L3) and pGEX-6P1-VP3(L4) were identified by digestion,PCR and sequencing,and transformed into E.coli Rosetta and BL21 cells.The target proteins of 32 kD,26 kD,30 kD and 31 kD were obtained by induction using IPTG at 1.0 mmol/L.The VP3(L1) and VP3(L4) protein reacted with IPNV positive serum in Western-blotting,indicating that the immunodominant region of the VP3 protein was located in 1~70 and 143~206 amino acid sequence of VP3 protein.The VP3 recombinant fusion protein can be used as the specific diagnosis antigen for ELISA assay.This study will serve as the basis for further precise identification of VP3 protein epitope. |
| |
| Keywords: | infectious pancreatic necrosis virus VP3 gene expression epitop |
| 本文献已被 CNKI 等数据库收录！ |
|
