表达传染性法氏囊病病毒VP2蛋白的转基因柔嫩艾美耳球虫的构建与鉴定
打开文本图片集
关键词：柔嫩艾美耳球虫；传染性法氏囊病病毒；VP2;mCherry;转基因中图分类号：S852.659.4;S852.723 文献标志码：A 文章编号：0366-6964（2026）02-1008-09
Construction and Identification of a Transgenic Eimeria tenella Expressing Infectious BursalDiseaseVirusVP2Protein
LIU Yuehui'， SUN Yingying²，WANG Haodi'， SUO Jingxia²， SONG Mingxin1*， LIU Xianyong² （1.Heilongjiang Key Laboratory for Zoonosis，Northeast Agricultural University，Harbin 15O03O，China; 2. College of Veterinary Medicine， China Agricultural University， Beijing lOo193， China）
Abstract： Infectious bursal disease （IBD） severely restricts the economic development of the poultry industry. Therefore，the development of safe and efficient IBD vaccines has become a key topic of considerable interest. To explore the feasibility of using Eimeria tenell as a live vector for the expression and delivery of infectious bursal disease virus （IBDV） antigens，this study constructed a genetically modified strain of E . tenella（Et-VP2） expressing the VP2 protein of the IBDV.An expression plasmid was constructed，digested with enzymes，and then transfected into E .tenella using nuclear transfection. Flow cytometry was utilized to sort and select positive strains expressing the red fluorescent reporter gene （mCherry），and the expression of the VP2 gene was identified at both the genetic and protein levels. Results showed that after three passages in vivo in chicks，the transgenic E .tenela expressing VP2 protein had a positive rate of fluorescence exceeding 95% ， confirming successful integration and stable expression of the foreign gene at both the genetic and protein levels （with fluorescence rates reaching 95% in the ofspring population）. Indirect immunofluorescence results indicated that the fusion protein （Mic2-linker-VP2） was localized at the apical end of the sporozoites of E . tenella. This study successfully established a stable transfection system for E . tenella，laying the foundation for subsequent research on the immune protection of genetically modified strains against infectious bursal disease.
Keywords： Eimeria tenella; infectious bursal disease virus; VP2;mCherry; transgenic ∗ Corresponding author： SONG Mingxin，E-mail： songmx@neau. edu. cn； LIU Xianyong，Email：liuxianyong@cau.edu.cn
传染性法氏囊病病毒（infectiousbursaldiseasevirus，IBDV）作为威胁全球养禽业的重要病原体，其VP2蛋白是诱导宿主免疫保护的关键抗原[1]当前商业化疫苗面临超强毒株免疫逃逸和母源抗体干扰等挑战，而基因工程疫苗如病毒活载体、亚单位疫苗和DNA疫苗虽各有优势，但仍存在表达效率或免疫原性不足的问题[2-3]。（剩余13937字）
