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,并誘導表達重組HSP70蛋白,試驗根據免疫蛋白組學研究的質譜結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(+),轉化Competent Rosetta2(DE3)p Lys S,通過IPTG誘導表達重組HSP70蛋白,并進行Western-blot分析。結果表明:HSP70序列全長為2 727 bp,最大ORF為2 010 bp。HSP70在誘導后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