摘要
Nebovirus(NeV)是导致犊牛腹泻的重要病原,是国内的新发病毒.VP1是NeV主要结构蛋白,在病毒的感染与免疫中发挥重要作用.以国内NeV流行毒株VP1基因序列为模板,进行密码子偏好性优化、序列合成并插入到pET28a(+)的Nde I和Xho I酶切位点间,构建了表达VP1完整编码框的pET28a-VP1-BL21(DE3)表达载体.SDS-PAGE结果显示,表达的目的蛋白大小约60 kDa,与预期相符;Western-blot结果显示该重组蛋白可以特异地与NeV抗血清反应;家兔免疫试验结果显示该重组蛋白可以刺激兔体产生特异性抗体.本试验成功表达了NeV VP1重组蛋白,具有良好的免疫原性.为进一步研究NeVVP1蛋白的功能和血清学检测方法奠定了基础.
Nebovirus(NeV) is an important pathogen causing diarrhea in calves and is an emerging causative agent in China. VP1 is the main structural protein of NeV and plays an important role in virus infection and immunity. Using the VP1 gene sequence of domestic epidemic NeV strain as template, the codon preference was optimized, the sequence was synthesized and inserted between Nde I and Xho I restriction sites of pET28 a(+), the complete VP1 coding frame pET28 a-VP1-BL21(DE3) expression vector was constructed. The results of SDS-PAGE showed that the size of the expressed VP1 protein was about 60 kDa, the Western-blot results showed that the recombinant protein could specifically react with NeV antiserum, and the rabbit immune test showed that it could stimulate the rabbit to produce specific antibodies. Recombinant NeV VP1 protein has been successfully expressed and has good immunogenicity, which lays a foundation for further study on the function and serological detection method of NeV VP1 protein.
作者
王珍贤
汤承
岳华
WANG Zhen-xian;TANG Cheng;YUE Hua(School of Animal Husbandry and Veterinary Medicine,Southwest Minzu University,Chengdu 610041,China)
出处
《西南民族大学学报（自然科学版）》
CAS
2021年第1期11-18,共8页
Journal of Southwest Minzu University(Natural Science Edition)
基金
西南民族大学新发动物疫病创新团队(2020NTD02)
西南民族大学研究生"创新型科研项目"(CX2017SZ061)
国家民委领军人才支持计划。
