中文摘要：

巨噬细胞炎症蛋白-3α是一种可诱导的分泌蛋白,通过相应受体对多种免疫细胞产生趋化,在多种疾病的免疫反应和炎症损伤中发挥重要作用。实验采用RT-PCR技术克隆出黄喉拟水龟巨噬细胞炎症蛋白-3α（MaCCL20）的成熟肽基因序列,通过双酶切将目的基因序列插入到表达载体pET-32a上,然后转化进大肠杆菌BL21中,经IPTG诱导后表达出了pET-32a-CCL20融合蛋白,以该融合蛋白为抗原制备多克隆抗体并进行Western-blotting鉴定及抑菌性检测。实验表明,融合蛋白在37℃、0.8 mmol/L IPTG,4 h条件下得到高效表达;经SDS-PAGE凝胶电泳和Western-blotting分析显示,所表达融合蛋白相对分子量为30 ku,与预测蛋白大小一致并与带His标签的多克隆抗体发生特异性反应。经His Bind镍柱纯化后SDS-PAGE电泳检测出现单一条带,说明获得了纯度较高的MaCCL20重组蛋白。抑菌实验表明,MaCCL20重组蛋白对金黄色葡萄球菌、粘质沙雷氏菌和嗜水气单胞菌都有较强的抑菌作用。

英文摘要：

Macrophage inflammatory protein-3α is a kind of inducible secreted protein which plays an important role in the immune response and inflammation damage in many diseases through the corresponding receptors on a variety of immune cells to produce chemokines.We obtained the ORF of MaCCL20 by RT-PCR,inserted into the expression vector pET-32a,and then transformed into E.coli BL21.We expressed the fusion protein pET-32a-CCL20 after IPTG induction,prepared the polyclonal antibodies and did research on Western blot and antimicrobial activity of it.The experiments show that the fusion protein is highly expressed in the conditions of 37 ℃,0.8 mmol/L IPTG for 4 hours.The SDS-PAGE gel electrophoresis figure shows that the relative molecular weight of the fusion protein is 30 ku which is the same as predicted before.We got a single band after His Bind nickel column purification indicating the high purity of MaCCL20 recombinant protein.The antibacterial experiments show that MaCCL20 recombinant protein has strong antibacterial effects on Staphylococcus aureus,Serratia marcescens,and Aeromonas hydrophila.