汉赛巴尔通体P26蛋白基因克

发布时间：2018-05-21 00:40

  本文选题：汉赛巴尔通体 + P蛋白基因　； 参考：《生物技术》2017年05期

【摘要】：[目的]研究汉赛巴尔通体(Bartonella henselae,Bh)P26蛋白的抗原特性。[方法]根据GenBank中Bh P26基因DNA序列设计特异性引物,对阳性猫血液样品进行PCR扩增,克隆测序后进行分子特征分析。构建表达载体pETP26,转化至感受态细胞E.coli BL21(DE3)进行诱导表达;将重组蛋白纯化后免疫小鼠,用间接ELISA抗体检测试剂盒检测抗体,分析其免疫学特性。[结果]P26基因全长738 bp,编码246个氨基酸。推导的P26蛋白氨基酸序列含有4个潜在的N-糖基化位点、4个酪蛋白激酶Ⅱ磷酸化位点、2个N-豆蔻酰化位点和1个信号肽序列;抗原表位主要集中在106~190位。SDS-PAGE和Western Blot分析结果表明,表达的分子量为35 kDa,可与B h阳性血清发生反应;重组蛋白免疫小鼠后其血清样本Bh抗体检测结果为阳性。[结论]P26重组蛋白在大肠杆菌的高效表达且具有良好的反应和免疫原性,为进一步探讨P26蛋白在Bh感染诊断研究中的潜在价值提供前期基础。
[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，

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