含有不同启动子、WPRE以及ITRs对重组杆状病毒表达新城疫F蛋白的影响  

Effects of Different Promoters, WPRE and ITRs on Recombinant Baculovirus Expression of Newcastle Disease F Protein

作　　者：曹德宸 叶磊 李维庆 李赛男 高冬妮 葛菁萍[1,2] Cao Dechen;Ye Lei;Li Weiqing;Li Sainan;Gao Dongni;Ge Jingping(Engineering Research Center of Agricultural Microbiology Technology,Ministry of Education,Heilongjiang University,Harbin 150500;Key Laboratory of Microbiology,College of Heilongjiang Province,School of Life Sciences,Heilongjiang University,Harbin 150080)

机构地区：[1]黑龙江大学农业微生物技术教育部工程研究中心,哈尔滨150500 [2]黑龙江大学生命科学学院黑龙江省普通高校微生物重点实验室,哈尔滨150080

出　　处：《中国农学通报》2020年第30期91-97,共7页Chinese Agricultural Science Bulletin

基　　金：黑龙江省自然科学基金项目“鸡传染性法氏囊病病毒基因工程疫苗的研制、教育部留学回国人员基金(鸡新城疫疫苗的研制)”(D0249);黑龙江省青年创新人才培养计划(UNPYSCT-2016178);黑龙江省科学基金(C2017056);黑龙江省博士后资助项目(LBH-Z17191)。

摘　　要：为了提高基因工程疫苗对新城疫病毒F蛋白的表达能力,从而为研制高效基因工程疫苗奠定了基础。利用Western blot技术检测含有不同启动子与表达元件的重组杆状病毒,在中国仓鼠卵巢细胞中其F蛋白的表达量的差异。Western blot结果显示,含有CMV立即早期增强子区与鸡肌动蛋白启动子区的复合启动子的质粒表达F蛋白量较含人巨细胞病毒立即早期启动子(CMV)的质粒提高了24.52%~231.18%,含有土拨鼠肝炎病毒转录后调控元件(WPRE)能够提高F蛋白表达量2.9~8.6倍,而腺伴随病毒末端反向重复序列(ITRs)对质粒F蛋白表达量提高1.95%~9.27%。CBA启动子的启动能力远高于CMV的启动能力,调控元件WPRE能够显著提高F蛋白的表达量,而ITRs元件对于F蛋白的表达量未产生显著提高。The aims are to improve the expression ability of genetically engineered vaccines to the Newcastle disease virus F protein, and lay a foundation for developing highly efficient and genetically engineered vaccines. The expression differences of baculovirus with different promoters and expression elements to F protein in Chinese hamster ovary cell were detected by Western blot. The results of Western blot showed that the expression level of complex promoter containing CMV enhancer region and chicken actin promoter region(CBA promoter) was higher than that of Cytomegaoviyns promoter(CMV) by 24.52%-231.18%, and the expression level of wood-chuck hepatitis virus post-transcriptional regulatory element(WPRE) was increased by 2.9-8.6 times, but adeno-associated virus inverted terminal repeats(ITRs) only increased the expression level of 1.95%-9.27%. In conclusion, the adjustment ability of CBA promoter is much higher than that of CMV promoter, while group WPRE could get a positive result, but group ITRs does not change significantly.

关 键 词：新城疫病毒 杆状病毒 土拨鼠肝炎病毒转录后调控元件 含有CMV立即早期增强子区与鸡肌动蛋白启动子区的复合启动子 

分 类 号：S852.65[农业科学—基础兽医学]