發(fā)布時(shí)間：2019-03-08 13:16

【摘要】：為了對(duì)DNA損傷誘導(dǎo)蛋白1(Ddi1-like)基因功能進(jìn)行研究,本研究參考GeneDB中豬帶絳蟲基因組注釋信息,通過(guò)設(shè)計(jì)5′和3′端特異性引物,利用cDNA末端快速擴(kuò)增法(RACE)獲得全長(zhǎng)cDNA,并在畢赤酵母GS115中進(jìn)行誘導(dǎo)表達(dá);將純化后的Ddi1-like重組蛋白免疫新西蘭大白兔,制備抗血清。結(jié)果,成功利用RACE獲得Ddi1-like基因的全長(zhǎng)cDNA序列;在真核表達(dá)系統(tǒng)中,絳蟲上的Ddi1-like重組蛋白獲得表達(dá);SDS-PAGE和Western-blot分析表明,獲得重組蛋白即為Ddi1-like蛋白;免疫后獲得抗血清,其效價(jià)達(dá)到1∶12 800。上述結(jié)果為后續(xù)研究Ddi1-like蛋白的酶切活性研究提供了基礎(chǔ)。
[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.
【作者單位】： 中國(guó)農(nóng)業(yè)科學(xué)院蘭州獸醫(yī)研究所家畜疫病病原生物學(xué)國(guó)家重點(diǎn)實(shí)驗(yàn)室農(nóng)業(yè)部獸醫(yī)公共衛(wèi)生重點(diǎn)開放實(shí)驗(yàn)室甘肅省動(dòng)物寄生蟲病重點(diǎn)實(shí)驗(yàn)室;
【基金】：國(guó)家重點(diǎn)基礎(chǔ)研究發(fā)展計(jì)劃(973)項(xiàng)目(2015CB150300)
【分類號(hào)】：S852.7
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本文編號(hào)：2436840