【摘要】 为表达捻转血矛线虫P450蛋白并分析其生物信息学特性,本研究利用RT-PCR扩增捻转血矛线虫P450基因,构建重组质粒p ET-P450,经双酶切鉴定,将其转化大肠杆菌后,采用SDS-PAGE检测P450蛋白的表达;利用生物信息学软件对P450蛋白的理化性质、结构特点进行分析。结果显示,正确构建了重组质粒p ET-P450且P450蛋白以包涵体的形式表达。生物信息学分析结果显示,P450蛋白分子式为C₁₁₅₄H₁₈₁₂N₃₁₄O₃₃₀S₁₅,为不稳定蛋白、无跨膜区、无信号肽,是一种含较多抗原表位的亲水性蛋白,有7个B细胞抗原表位和11个T细胞抗原表位,有望用作捻转血矛线虫耐药性的诊断抗原。本研究获得了P450重组蛋白,为捻转血矛线虫耐药性快速检测方法和新药开发奠定物质基础。更多还原

【Abstract】 To analyze the bioinformatics characteristics of the P450 gene of Haemonchus contortus and obtain the P450recombinant protein,RT-PCR technology was used to amplify the P450 gene of Haemonchus contortus.A p ET-P450 recombinant expression vector was constructed and identified by double enzyme digestion,and later transformed into E.coli and induced for protein expression.The results showed that the recombinant plasmid p ET-P450 was correctly constructed and P450 protein is expressed as inclusion body.The physicochemical properties and structural characteristics of P450 protein were analyzed by bioinformatics software.Bioinformatics analysis results showed that the molecular formula of P450 protein is C₁₁₅₄H₁₈₁₂N₃₁₄O₃₃₀S₁₅.The P450 protein is an unstable protein with no transmembrane region and no signal peptide region.P450 protein is a kind of hydrophilic protein with 7 B-cell epitopes and 11 T-cell epitopes,thus it is expected to be used as a diagnostic antigen for resistance to Haemonchus contortus.The recombinant P450 protein was successfully purified in this study,providing a theoretical basis for the rapid detection of resistance to Haemonchus contortus and the development of new drugs.更多还原