本文選題：斯氏艾美耳球蟲 + 熱休克蛋白(HSP)　； 參考：《黑龍江畜牧獸醫(yī)》2017年07期

【摘要】：為了獲得斯氏艾美耳球蟲(E.stiedai)的熱休克蛋白70(HSP70)的全基因序列,并誘導(dǎo)表達重組HSP70蛋白,試驗根據(jù)免疫蛋白組學(xué)研究的質(zhì)譜結(jié)果獲得的部分氨基酸序列和已發(fā)布近源物種HSP70蛋白序列設(shè)計引物,通過c DNA末端快速擴增法(RACE)獲得E.stiedai HSP70的全基因序列,將其連入原核表達載體p ET-28a(+),轉(zhuǎn)化Competent Rosetta2(DE3)p Lys S,通過IPTG誘導(dǎo)表達重組HSP70蛋白,并進行Western-blot分析。結(jié)果表明:HSP70序列全長為2 727 bp,最大ORF為2 010 bp。HSP70在誘導(dǎo)后37℃培養(yǎng)4 h條件下有表達,但全部為不溶性表達;在誘導(dǎo)后20℃過夜培養(yǎng)條件下無表達。說明試驗成功獲得E.stiedai HSP70的全基因序列以及重組HSP70蛋白。
[Abstract]:In order to obtain the whole gene sequence of heat shock protein 70 (HSP70) of E. stiedai, and to induce the expression of recombinant HSP70 protein, A primer was designed based on the results of mass spectrometry and HSP70 protein sequence of the published near-source species. The whole gene sequence of E.stiedai HSP70 was obtained by rapid amplification of c DNA terminal. The recombinant HSP70 protein was transfected into prokaryotic expression vector pET-28a (pET-28a) and transformed into Competent Rosetta2(DE3)p Lys S.The recombinant HSP70 protein was induced by IPTG and analyzed by Western-blot. The results showed that the full length of the 1: HSP70 sequence was 2 727 BP, and the maximum ORF was 2 010 bp.HSP70, which was expressed at 37 鈩，

本文編號：1986355