猪带绦虫Ddi1-like蛋白的真核表达及其抗血清的制备
发布时间：2019-03-08 13:16
【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。
[Abstract]:In order to study the function of DNA damage inducible protein 1 (Ddi1-like) gene, we designed 5 'and 3' end-specific primers according to the genomic annotation information of Taenia solium in GeneDB. The full-length cDNA, was obtained by rapid amplification of cDNA terminal (RACE) and expressed in Pichia pastoris GS115. Anti-serum was prepared by immunizing New Zealand rabbits with purified Ddi1-like recombinant protein. Results: the full-length cDNA sequence of Ddi1-like gene was successfully obtained by RACE, the recombinant Ddi1-like protein was expressed in the eukaryotic expression system, and the SDS-PAGE and Western-blot analysis showed that the recombinant protein was Ddi1-like protein. The antiserum was obtained after immunization, and the titer was up to 1:12 800. These results provide a basis for further study on the enzyme digestion activity of Ddi1-like protein.
【作者单位】： 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;
【基金】：国家重点基础研究发展计划(973)项目(2015CB150300)
【分类号】：S852.7
，
本文编号：2436840
[Abstract]:In order to study the function of DNA damage inducible protein 1 (Ddi1-like) gene, we designed 5 'and 3' end-specific primers according to the genomic annotation information of Taenia solium in GeneDB. The full-length cDNA, was obtained by rapid amplification of cDNA terminal (RACE) and expressed in Pichia pastoris GS115. Anti-serum was prepared by immunizing New Zealand rabbits with purified Ddi1-like recombinant protein. Results: the full-length cDNA sequence of Ddi1-like gene was successfully obtained by RACE, the recombinant Ddi1-like protein was expressed in the eukaryotic expression system, and the SDS-PAGE and Western-blot analysis showed that the recombinant protein was Ddi1-like protein. The antiserum was obtained after immunization, and the titer was up to 1:12 800. These results provide a basis for further study on the enzyme digestion activity of Ddi1-like protein.
【作者单位】： 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;
【基金】：国家重点基础研究发展计划(973)项目(2015CB150300)
【分类号】：S852.7
，
本文编号：2436840
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