摘要
以防御素为研究对象探讨其药用开发价值已是国内外生物医学研究领域的热点之一,鸡β-防御素-1(Gallinacin-1,Gal-1)对许多致病菌都具有杀菌作用,具有很好的研究开发价值。本研究以含有Gal-1成熟肽全长cDNA的重组质粒pMD19-T-Gal-1为模板,通过PCR在Gal-1成熟肽基因C末端引入6×His-tag,并将其克隆到pcDNA3.1(+)载体中,构建真核表达质粒pcDNA3.1(+)-Gal-1-His;经脂质体介导转染Flp-In-293细胞,48h后间接免疫荧光分析,结果重组质粒pcDNA3.1(+)-Gal-1-His转染的Flp-In-293细胞中有绿色荧光,而阴性对照中无荧光,表明Gal-1-His在Flp-In-293细胞中得到了表达。本研究结果将为进一步研究Gal-1的功能奠定基础。
To defensins as a template or directly discussing the development of the medicinal value of defensins have been one of the hot areas of biomedical research at home and abroad.Chicken beta-defensin-1（Gallinacin-1,Gal-1） has antimicrobial activity against potentially pathogens and is worth being investigated and developed.The core coding regions sequence of Gal-1 fused with 6×His in the C-terminal was amplified by PCR from pMD19-T-Gal-1 and the Gal-1-His gene was inserted into the eukaryotic expression plasmid pcDNA3.1（＋）,then the recombinant pcDNA3.1（＋）-Gal-1-His plasmids were transfected into Flp-In-293 cells using lipofectamine reagent and indirect immunofluorescence was used to analysis the results after transfection 48 hours.The fusion protein Gal-1-His was successfully expressed in Flp-In-293 cells by the detection of bright green fluorescence.This study provides a basis for further investigation of Gal-1 function.
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2011年第5期539-543,共5页
Genomics and Applied Biology
基金
农业行业专项研究计划(nxhyzx07-039)
国家高技术研究发展计划(863)(2011AA100301)
江苏省家禽科学研究所青年基金项目(S030)共同资助
关键词
鸡
β-防御素-1
真核表达
Chicken
Beta-defensin-1
Eukaryotic expression
作者简介
通讯作者，lhfxy_002@yahoo．com．cn
