中文摘要：

【目的】克隆布鲁氏菌Ⅳ型分泌系统效应分子BPE123和BPE275的基因并进行诱导表达。【方法】从牛种布鲁氏菌2308株基因组中PCR扩增BPE123和BPE275基因片段,亚克隆到pGEM-T Easy载体中并测序。将BPE123和BPE275基因克隆至融合表达载体pET-28a,构建表达重组质粒pET-28a-BPE123和pET-28aBPE275,在大肠杆菌中进行诱导表达,Western blot分析重组蛋白His-BPE123和His-BPE275的免疫学特性。【结果】PCR扩增获得了462bp的BPE123基因片段和762bp的BPE275基因片段,成功构建了pET-28a-BPE123、pET-28a-BPE275原核表达载体,并在大肠杆菌中成功表达了BPE123和BPE275蛋白,经SDS-PAGE检测,重组融合蛋白BPE123、BPE275的分子质量分别约为22和31ku,布鲁氏菌免疫动物血清能特异性识别表达的蛋白。【结论】克隆了分泌蛋白BPE123、BPE275基因片段,并成功表达了布鲁氏菌Ⅳ型分泌系统效应分子BPE123和BPE275。

英文摘要：

【Objective】This study aimed to clone and express effector molecules BPE123 and BPE275of BrucellatypeⅣ secretion system.【Method】The BPE123 and BPE275genes were obtained fromBrucella abortus 2308 genome.Then,they were linked to the vector pGEM-T Easy and cloned to expression vector pET-28 ato construct recombinant plasmids pET-28a-BPE123 and pET-28a-BPE275.These constructs were expressed on E.coli BL21 cells,and the immunological characteristics were analyzed using Western blot analysis.【Result】Full lengths of BPE123 and BPE275were 462 bp and 762 bp.Prokaryotic expression vector pET-28a-BPE123 and pET-28a-BPE275 were constructed.The BPE123 and BPE275protein were successfully expressed in E.coli.SDS-PAGE showed proteins with molecular weights of 22 ku and 31 ku.The expressed proteins can be specifically identified by immune serum against Brucella.【Conclusion】This study successfully cloned BPE123 and BPE275genes of Brucella abortus 2308 and expressedthem.