生物技术布鲁氏菌Ompbls融合基因重组表达载体的构建与诱导表达.doc

内蒙古科技大学 本科生毕业论文 题 目：布鲁氏菌Omp31-bls融合基因重组表达载体的构建与诱导表达 学生姓名：xxxxx 学 号：xxxxx 专 业：生物技术 班 级：生技 2008 指导教师：xxx 布鲁氏菌Omp31-bls融合基因重组表达载体的构建与诱导表达 摘 要 布鲁氏菌病（Brucellosis）是当今在全世界广泛流行的一种人畜共患的传染疾病，该病是由属于布鲁氏菌属的一种小型革兰氏阴性菌引起的。布鲁氏菌（Brucella）能够直接寄生到人和动物的细胞内（特别是生殖系统细胞），主要引起死胎、流产、不孕等症状。近几年来，疫情反弹，死灰复燃，给畜牧业和人类健康带来及其严重的危害。由此可见，布鲁氏菌病严重威胁着我国经济建设、人民健康、社会稳定以及国家的安全。预防并且控制甚至根除布鲁氏菌病已经成为人们的当务之急。 本实验以非常重要的两种蛋白OMP31和BLS作为研究对象，利用融合PCR技术，将Omp31和bls基因进行融合重组，随后构建Omp31-bls融合基因的原核重组表达载体pET-Omp31-bls。转化表达型宿主菌TL129感受态细胞之后，利用IPTG对融合基因进行诱导表达。结果显示pET-Omp31-bls原核重组表达载体构建正确，rOMP31-BLS融合蛋白被成功表达。 pET-28a载体 The Construction and Induced Expression of the Recombinant Expression Plasmid pET-Omp31-bls Abstract Brucellosis is a zoonosis nowadays spreading all over the world. It is caused by a small bacterium called Brucella which is a kind of Gram-negative bacterium. The Brucella could directly parasitize in human and animal cells (especially in the reproductive system cells), and the symptoms of Brucellosis are fetal death, abortion and sterility of livestock. In the recent years epidemic situation brings serious harm to the development of animal husbandry and human health. Therefore, Brucellosis is a severe threat to the economy construction, public health and the national security to our courtry. Prevention, control and even eradication of brucellosis are becoming the most pressing matters of the moment. In this study, OMP31 and BLS, which are very important proteins in B. suis, were taken as our reseach subjects. Omp31 was fused with bls by the technology of the fusion PCR, and then constructed a recombinant expression plasmid pET-Omp31-bls carrying the fusion gene Omp31-bls. After transferring into the competent cells of the expression strain TL129, the fusion gene was induced with IPTG. The results of the expriment showed that the recombinant expression plasmid pET-Omp31-bls was correctly constructed, and the recombinant protein rOMP31-BLS was successfully expressed. Key words: Brucella; B