机构地区: 扬州大学兽医学院江苏省人兽共患病学重点实验室
出 处: 《中国家禽》 2013年第18期19-22,共4页
摘 要: 为了获得具有良好生物学活性且具有较高表达量的鸡白细胞介素-17(ChIL-17),本研究在前期研究工作基础上,将ChIL-17的编码基因按照真核细胞(昆虫细胞)偏爱的密码子进行优化改造,经全基因合成后插入到转座载体pFastBacTMⅠ中,构建重组转移质粒pfast-mod.ChIL-17并转化DH10Bac感受态细胞。通过位点特异性转座将mod.ChIL-17基因整合到穿梭质粒Bacmid中,获得重组穿梭质粒Bacmid-mod.ChIL-17。应用脂质体将重组穿梭质粒转染Sf9昆虫细胞,获得重组杆状病毒rBac-mod.ChIL-17。重组病毒传代扩增感染Sf9细胞,通过间接免疫荧光检测目的蛋白的表达。结果表明,经过优化的ChIL-17在杆状病毒系统中获得表达。 In order to improve the expression of chicken interleukin-17 (ChIL-17)and obtain good biologic activity protein, the whole length of ChIL-17 gene in accordance with codon preference of eukaryotic cells (insect cells) were optimized and synthesized. The mod.ChIL-17 was cloned into transfer vector pFastBacTM I , and transformated into DH10Bac competent E.coli. By site-specific transposition, mod.ChIL-17 gene was integrated into shuttle vector Bacmid, and transfeeted into Sf9 insect cells mediated by lipofectine to produce recombinant baculovirus.Indirect immunofluorescenee assay confirmed that mod.ChIL-17 was expressed successfully in baculovirus expression system.
领 域: [生物学]