Construction and expression efficiency of the recombinant plasmid pBI-Eg95 of Echinococcus granulosus in the agrobacterium tumefaciens LBA4404 strain

ZHOU Hui; et al

您当前的位置：

首页 >

文章列表页 >

Construction and expression efficiency of the recombinant plasmid pBI-Eg95 of Echinococcus granulosus in the agrobacterium tumefaciens LBA4404 strain

更新时间：2024-01-17

- Construction and expression efficiency of the recombinant plasmid pBI-Eg95 of Echinococcus granulosus in the agrobacterium tumefaciens LBA4404 strain
- Journal of Chongqing Medical University Vol. 34, Issue 8, Pages: 977-980(2009)
- 作者机构：
  
  重庆医科大学附属第一医院传染病寄生虫病研究所
- 作者简介：
- 基金信息：
- DOI：
  CLC： R392
- Published：2009，
扫描看全文
[1]周辉,李文桂,叶艳菊.细粒棘球绦虫重组质粒pBI-Eg95构建及其在根癌农杆菌LBA4404中表达效率的研究[J].重庆医科大学学报,2009,34(08):977-980.

ZHOU Hui, et al. Construction and expression efficiency of the recombinant plasmid pBI-Eg95 of Echinococcus granulosus in the agrobacterium tumefaciens LBA4404 strain. [J]. Journal of Chongqing Medical University 34(8):977-980(2009)
[1]周辉,李文桂,叶艳菊.细粒棘球绦虫重组质粒pBI-Eg95构建及其在根癌农杆菌LBA4404中表达效率的研究[J].重庆医科大学学报,2009,34(08):977-980. DOI：

ZHOU Hui, et al. Construction and expression efficiency of the recombinant plasmid pBI-Eg95 of Echinococcus granulosus in the agrobacterium tumefaciens LBA4404 strain. [J]. Journal of Chongqing Medical University 34(8):977-980(2009) DOI：

摘要

目的:构建细粒棘球绦虫重组质粒pBI-Eg95

分析其在根癌农杆菌（Agrobacterium tumefaciens

At）LBA4404株中的表达效率。方法:从细粒棘球蚴包囊中分离原头节

超声粉碎后抽提总RNA

采用RT-PCR方法扩增Eg95编码基因

将该基因定向克隆到植物表达载体pBI121中构建pBI-Eg95重组质粒;电穿孔转化A（tLBA4404）

抽提质粒进行酶切及PCR鉴定。用IPTG分别诱导rAt0、1、3、5、7、9、11和13h

用SDS-PAGE和Western blot鉴定所表达的蛋白质。结果:471bpEg95基因克隆成功。经酶切及PCR证实重组质粒pBI-Eg95成功转入A（tLBA4404）。用Bio-Rad Quantity one分析系统分析

表达产物分子质量（Mr）约为16.5kD

且能被感染细粒棘球蚴的鼠血清特异识别

表达效率为约占LBA4404菌体总蛋白的20%。结论:成功构建了细粒棘球绦虫重组质粒pBI-Eg95

且能在A（tLBA4404）中表达

为进一步研究细粒棘球绦虫转基因植物疫苗奠定了基础。

Abstract

Objective:To construct the recombinant plasmid pBI-Eg95 of E.granulosus

and to study its expression efficiency in the Agrobacterium tumefaciens(At) LBA4404 strain.Methods:Total RNA was extracted from hydatid cyst protoscoleces shattered by supersound.The Eg95 coding gene was amplified by RT-PCR

and then was cloned into the plant expression vector pBI121 to construct the recombinant plasmid pBI-Eg95.The recombinant plasmid was electroporated into At(LBA4404) strain.The recombinant At(rAt) was induced by IPTG for 0

11or13 hour

respectively.The expression product was identified by SDS-PAGE and Western blot assay.Results:The 471bp Eg95 coding gene was successfully amplified

and the recombinant pBI-Eg95 plasmid was successfully constructed

which was confirmed by restriction endonuclease digestion and PCR.The recombinant plasmid pBI-Eg95 was expressed successfully in the rAt

and the expression product with an approximately Mr of 16.5 kD could be detected by sera from mice infected by E.granulosus

and the expression efficiency was about 20 percent of the total bacterial protein.Conclusion:The recombinant plasmid could provide the basis for further research of the transgenic plant vaccine of E.granulosus.

关键词

细粒棘球绦虫重组质粒pBI-Eg95根癌农杆菌构建表达效率

Keywords

Echinococcus granulosusRecombinant pBI-Eg95 plasmidAgrobacterium tumefaciensconstructionExpression efficiency

references

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Construction and identification of the recombinant pCD-Eg95 plasmid of Echinococcus granulosus

The construction and significance of maspin/PCR2.1 expression vector

Construction and confirmation of tetracycline-dependent regulatory system in one vector

Construction of human single-chain antibody gene for breast cancer

Dynamic observation on IgG,its subclass and IgE in sera of mice immunized by recombinant BCG-Eg95 vaccine of Echinococcus granulosus

Related Author

et al

ZHOU Hui

et al

YAN Xiong

王蜀强

ZHOU Zhi

et al

雷宇

Related Institution

重庆医科大学临床学院普外科

Department of General Surgery,College of Clinical Medicine,Chongqing Medical University

重庆医科大学附属第二医院肝病治疗研究中心

Department of Diseases Therapy Center,the Second Affiliated Hospital,Chongqing Medical Univesity

重庆医科大学基础医学院核医学教研室

⁰