CAJ | 학술논문

　　为了使鸭坦布苏病毒（Duck tembusu virus, DTMUV）的 E 蛋白在 Bac-to-Bac 杆状病毒/昆虫细胞表达系统中高效表达,本研究将 DTMUV 的 E 基因按照昆虫细胞偏嗜性密码子进行了优化，合成了基因 optiE，并将其插入到 pFastBac HT 载体上，得到质粒 pFastBac-optiE。将该质粒转化 DH10BAC 感受态细胞，经抗性和蓝白斑筛选，获得重组转座子 rBacmid-optiE。将rBacmid-optiE 转染对数生长期的 Sf9昆虫细胞，间接免疫荧光试验和 Western blot 方法都证实目的蛋白在重组杆状病毒感染的Sf9昆虫细胞中获得了表达。本研究为进一步研究 DTMUV E 蛋白的结构与功能奠定了基础。
　　위료사압탄포소병독（Duck tembusu virus, DTMUV）적 E 단백재 Bac-to-Bac 간상병독/곤충세포표체계통중고효표체,본연구장 DTMUV 적 E 기인안조곤충세포편기성밀마자진행료우화，합성료기인 optiE，병장기삽입도 pFastBac HT 재체상，득도질립 pFastBac-optiE。장해질립전화 DH10BAC 감수태세포，경항성화람백반사선，획득중조전좌자 rBacmid-optiE。장rBacmid-optiE 전염대수생장기적 Sf9곤충세포，간접면역형광시험화 Western blot 방법도증실목적단백재중조간상병독감염적Sf9곤충세포중획득료표체。본연구위진일보연구 DTMUV E 단백적결구여공능전정료기출。
To express E protein of Duck tembusu virus (DTMUV), E gene was optimized according to the bias codon usages of insect cells. The optimized E gene (opti-E) was cloned into the multiple cloning site of pFastBacHT vector of the Bac-to-Bac expression system. The recombinant plasmid pFastBac-optiE was then transformed into DH10Bac for blue/white selection and antibiotics selection. The positive recombinant bacmid rBacmid-optiE was transfected to Sf9 in logarithmic growth phase for harvesting recombinant baculovirus. The E protein expressed in Sf9 was confirmed in Western blot and IFA. The expression of E protein optimized was based for the further study of its structure and its function.