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布鲁氏菌病是一种人畜共患病,严重威胁着人和家畜的健康.Omp31与BLS都是布鲁氏菌重要的表面抗原基因,将重组Omp31分子与BLS分子融合表达之后,能够进一步加强抗原蛋白Omp31的免疫原性.本研究以布鲁氏菌融合基因Omp3148-74-BLS作为研究对象,双酶切后克隆至含有EGFP基因的pJG053植物表达载体上.之后将重组质粒转化到农杆菌GV3101中,利用该农杆菌侵染烟草,通过观察EGFP基因的表达来判断融合基因Omp314-74-BLS的表达,再提取侵染烟草叶片总RNA后反转录cDNA,进行RT-PCR来检测融合基因是否表达.结果显示,融合蛋白Omp3148-74-BLS顺利在烟草中表达.“,”Brucellosis is a zoonosis,seriously threatening the health of people and livestock.Omp31 and BLS are important brucella surface antigen.Fusion protein of Omp31 and BLS is expected to enhance the immunogenicity of Omp31 protein.This project aims on plant expression of fusion gene in tobacco as part of plant vaccine construction.Towards that end,brucella fusion gene Omp3148-74-BLS is cloned into plant expressing vector pJG053 in frame of EGFP gene that serves as a reporter of fusion gene expression.Then the recombinant plasmid is transformed into bacterium agrobacterium GV3101 that is used to infect tobacco.The expression of fusion gene Omp3148-74-BLS is demonstrated by observing the EGFP gene expression with fluorescence microscopy.The fusion gene expression is further confirmed by RT-PCR for total RNA extracted from tobacco leafs that are infected by recombinant agrobacterium GV3101.Results show that the fusion protein Omp3148-74-BLS is successfully expressed in tobacco.