旋毛虫T626-55基因减毒沙门氏菌疫苗的构建及其在小鼠体内表达  

Construction and in vivo Expression of Trichinella spralis T626-55 Gene Vaccine Carried by Attenuated Salmonella

作　　者：杜婧[1] 刘攀[1] 唐斌[1] 高鹤[1] 刘明远[1] 

机构地区：[1]吉林大学人兽共患病研究所,长春130062

出　　处：《吉林农业大学学报》2011年第6期682-686,共5页Journal of Jilin Agricultural University

基　　金：国家自然科学基金项目(NSFC30771885);国家杰出青年基金项目(NSFC30825033)

摘　　要：采用PCR方法扩增旋毛虫强反应原性基因T626-55,与真核表达载体pcDNA3.1相连,构建携带编码旋毛虫T626-55基因真核表达质粒的沙门氏菌,将构建好的重组质粒T626-55-pcDNA3.1通过电穿孔转化入沙门菌中。小鼠口服免疫该重组沙门菌,5 d后通过反转录PCR(RT-PCR)和间接免疫荧光(IFA)检测目的基因在小鼠组织内的转录和表达情况。结果表明:成功构建含有目的基因真核表达质粒的沙门菌T626-55-pcD-NA3.1-PQ,通过口服免疫该菌,在小鼠脾脏及派伊尔氏结(PP)内检测到目的基因的转录,在脾脏内检测到目的基因表达。Highly immunogenic gene T626-55 of TrichineUa spiralis was amplified using PCR reaction and cloned into pcDNA3.1 eukaryotic expression plasmid. The recombinant plasmid was transformed into S. enterica Typhimurium phoP/phoQ strain by electroporation. In vivo expression of the target gene was testified. Mice were orally administrated with the recombinant bacteria and after 5 days of administration, spleen and Peyer＇s patch were aseptically removed for reverse transcription-polymerase chain reaction （RT- PCR） and indirect fluorescence assay （IFA）. The results show that Salmonella harboring T626-55 gene coding eukaryotic plasmid T626-55-pcDNA3.1-PQ was successfully constructed. The transcription of T626-55 gene was detected in spleen and pp, and expression of T626-55 protein was confirmed in spleen.

关 键 词：旋毛虫 T626-55基因 沙门氏菌 反转录PCR 间接免疫荧光 

分 类 号：S852.7[农业科学—基础兽医学]