猪带绦虫Ddi1-like蛋白的真核表达及其抗血清的制备  

Eukaryotic expression and anti-serum preparation of Ddi1-like protein from Taenia solium

作　　者：何伟[1] 殷静[1] 岳生赟 刘仲藜[1] 王帅[1] 才学鹏[1] 

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部兽医公共卫生重点开放实验室甘肃省动物寄生虫病重点实验室,甘肃兰州730046

出　　处：《中国兽医科学》2017年第5期582-586,共5页Chinese Veterinary Science

基　　金：国家重点基础研究发展计划(973)项目(2015CB150300)

摘　　要：为了对DNA损伤诱导蛋白1(Ddi1-like)基因功能进行研究,本研究参考GeneDB中猪带绦虫基因组注释信息,通过设计5′和3′端特异性引物,利用cDNA末端快速扩增法(RACE)获得全长cDNA,并在毕赤酵母GS115中进行诱导表达;将纯化后的Ddi1-like重组蛋白免疫新西兰大白兔,制备抗血清。结果,成功利用RACE获得Ddi1-like基因的全长cDNA序列;在真核表达系统中,绦虫上的Ddi1-like重组蛋白获得表达;SDS-PAGE和Western-blot分析表明,获得重组蛋白即为Ddi1-like蛋白;免疫后获得抗血清,其效价达到1∶12 800。上述结果为后续研究Ddi1-like蛋白的酶切活性研究提供了基础。To functionally study the DNA damage-inducible-like （Ddil-like） protein from Teania solium,according to gene annotations of K solum genome in GeneDB,the cDNA of Ddil-like gene was ampli- fied by RACE （rapid-amplification of cDNA ends） technique using the specific 5＇and 3＇primers,and was then cloned into vecter pPIC9K. Then the recombinant protein was induced in E pastoris. New Zealand white rabbits were immuned by the purified recombinant Ddil-like protein to produce the anti-serum. In result,the full ORF of Ddil-like gene was successfully obtained. The recombinant Ddil-like protein was produced in^pastoris expression system. The recombinant protein expression was confirmed by SDS-PAGE and Western-blot analysis. The antibody titer of the anti-serum could reach to 1：12 800. The above-men- tioned results lay a foundation to further studies on the proteolytic activity of the Ddil-like protein.

关 键 词：绦虫 Ddi1-like蛋白 真核表达 蛋白酶 

分 类 号：S852.734[农业科学—基础兽医学]