本文選題：漢賽巴爾通體 + P蛋白基因�。� 參考：《生物技術(shù)》2017年05期

【摘要】：[目的]研究漢賽巴爾通體(Bartonella henselae,Bh)P26蛋白的抗原特性。[方法]根據(jù)GenBank中Bh P26基因DNA序列設(shè)計特異性引物,對陽性貓血液樣品進行PCR擴增,克隆測序后進行分子特征分析。構(gòu)建表達載體pETP26,轉(zhuǎn)化至感受態(tài)細胞E.coli BL21(DE3)進行誘導(dǎo)表達;將重組蛋白純化后免疫小鼠,用間接ELISA抗體檢測試劑盒檢測抗體,分析其免疫學特性。[結(jié)果]P26基因全長738 bp,編碼246個氨基酸。推導(dǎo)的P26蛋白氨基酸序列含有4個潛在的N-糖基化位點、4個酪蛋白激酶Ⅱ磷酸化位點、2個N-豆蔻酰化位點和1個信號肽序列;抗原表位主要集中在106~190位。SDS-PAGE和Western Blot分析結(jié)果表明,表達的分子量為35 kDa,可與B h陽性血清發(fā)生反應(yīng);重組蛋白免疫小鼠后其血清樣本Bh抗體檢測結(jié)果為陽性。[結(jié)論]P26重組蛋白在大腸桿菌的高效表達且具有良好的反應(yīng)和免疫原性,為進一步探討P26蛋白在Bh感染診斷研究中的潛在價值提供前期基礎(chǔ)。
[Abstract]:[objective] to study the antigenic characteristics of BhmP 26 protein from Bartonella henselaeae of Bartonella Henselaeae. [methods] according to the DNA sequence of BhP26 gene in GenBank, the specific primers were designed. The positive cat blood samples were amplified by PCR and cloned and sequenced. The expression vector pETP26 was constructed and transformed into E.coli BL21DE3 to induce the expression, and the recombinant protein was purified and immunized to mice. The antibody was detected by indirect ELISA antibody detection kit, and its immunological characteristics were analyzed. [results] the total length of P26 gene was 738 BP, encoding 246 amino acids. The deduced amino acid sequence of P26 protein contained four potential N-glycosylation sites, four casein kinase 鈪，

本文編號：1916982