Construction of transgenic Eimeria tenella over-expressing ApiAP2 transcription factor
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- 英文论文题名：Construction of transgenic Eimeria tenella over-expressing ApiAP2 transcription factor
- 论文作者：Dandan Hu ; Guangping Huang ; Chunhui Duan ; Jingxia Suo ; Xianyong Liu ; Xun Suo
- 英文论文作者：Dandan Hu ; Guangping Huang ; Chunhui Duan ; Jingxia Suo ; Xianyong Liu ; Xun Suo ; National Animal Protozoa Laboratory ; College of Veterinary Medicine ; China Agricultural University
- 年：2016
- 作者机构：National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University;
- 英文论文关键词：Eimeria tenella ; apicomplexan AP2 ; over-expressing
- 会议召开时间：2016-08-08
- 会议录名称：中国动物学会寄生虫学专业委员会第十届全国寄生虫学青年工作者学术研讨会论文摘要集
- 英文会议录名称：The Abstracts of the Tenth Symposium for Yong Scientists of China Society of Parasitology
- 语种：英文
- 分类号：S852.723
- 学会代码：JISC
- 会议名称：中国动物学会寄生虫学专业委员会第十届全国寄生虫学青年工作者学术研讨会
- 会议地点：中国四川成都
- 主办单位：中国动物学会寄生虫学专业青年委员会
- 学会名称：中国动物学会寄生虫学专业委员会
- 页数：1
- 文件大小：152k
- 原文格式：D
- 会议级别：全国
摘要
Objective: Chicken coccidiosis, caused by Eimeria sp., leads to huge economic losses in the poultry industry. This disease could be controlled by vaccination with different Eimeria precocious lines, which was selected by multiple continued propagation in chicken. Recent researches on Plasmodium revealed that a transcription factor apicomplexanAP2(ApiAP2) played a crucial role in the sexual commitment in its lifecycle. Here, to characterize the orthologous ApiAP2 gene in E. tenella(EtAp2) and its potential role in regulating Eimeria precocious, we constructed a transgenic E. tenella line over-expressing EtAp2. Methods: The transfection plasmid consists of a pyrimethamine selection maker(Toxoplasma gondii dihydrofolate reductase-thymidylate synthase) and fused immediately with an enhanced YFP reporter gene, the complete EtAp2 coding sequence was then linked by a "self-cleaving" 2A peptide sequence. This plasmid was regulated by the E. tenella Histone 4 promoter and C-terminal untranslated region of E. tenella Actin. Purified E. tenella sporozoites were transfected with the over-expressing vector and then inoculated into the MDBK bovine kidney cells and chickens. Results: The total transfection rate was 0.07% in the first generation oocysts measured by flow cytometry analysis. Both the transiently transfected sporozoites and the stable transfected E. tenella-Ap2 oocysts were expressing e YFP in a nuclear expressing manner. Conclusion: We constructed a transgenic E. tenella line over-expressing EtAp2 successfully. Our results established a basis for the further study on mining the downstream genes regulated by this transcription factor, which might helpful in understanding the mechanism of precocious.
Objective: Chicken coccidiosis, caused by Eimeria sp., leads to huge economic losses in the poultry industry. This disease could be controlled by vaccination with different Eimeria precocious lines, which was selected by multiple continued propagation in chicken. Recent researches on Plasmodium revealed that a transcription factor apicomplexanAP2(ApiAP2) played a crucial role in the sexual commitment in its lifecycle. Here, to characterize the orthologous ApiAP2 gene in E. tenella(EtAp2) and its potential role in regulating Eimeria precocious, we constructed a transgenic E. tenella line over-expressing EtAp2. Methods: The transfection plasmid consists of a pyrimethamine selection maker(Toxoplasma gondii dihydrofolate reductase-thymidylate synthase) and fused immediately with an enhanced YFP reporter gene, the complete EtAp2 coding sequence was then linked by a "self-cleaving" 2A peptide sequence. This plasmid was regulated by the E. tenella Histone 4 promoter and C-terminal untranslated region of E. tenella Actin. Purified E. tenella sporozoites were transfected with the over-expressing vector and then inoculated into the MDBK bovine kidney cells and chickens. Results: The total transfection rate was 0.07% in the first generation oocysts measured by flow cytometry analysis. Both the transiently transfected sporozoites and the stable transfected E. tenella-Ap2 oocysts were expressing e YFP in a nuclear expressing manner. Conclusion: We constructed a transgenic E. tenella line over-expressing EtAp2 successfully. Our results established a basis for the further study on mining the downstream genes regulated by this transcription factor, which might helpful in understanding the mechanism of precocious.
Objective: Chicken coccidiosis, caused by Eimeria sp., leads to huge economic losses in the poultry industry. This disease could be controlled by vaccination with different Eimeria precocious lines, which was selected by multiple continued propagation in chicken. Recent researches on Plasmodium revealed that a transcription factor apicomplexanAP2(ApiAP2) played a crucial role in the sexual commitment in its lifecycle. Here, to characterize the orthologous ApiAP2 gene in E. tenella(EtAp2) and its potential role in regulating Eimeria precocious, we constructed a transgenic E. tenella line over-expressing EtAp2. Methods: The transfection plasmid consists of a pyrimethamine selection maker(Toxoplasma gondii dihydrofolate reductase-thymidylate synthase) and fused immediately with an enhanced YFP reporter gene, the complete EtAp2 coding sequence was then linked by a "self-cleaving" 2A peptide sequence. This plasmid was regulated by the E. tenella Histone 4 promoter and C-terminal untranslated region of E. tenella Actin. Purified E. tenella sporozoites were transfected with the over-expressing vector and then inoculated into the MDBK bovine kidney cells and chickens. Results: The total transfection rate was 0.07% in the first generation oocysts measured by flow cytometry analysis. Both the transiently transfected sporozoites and the stable transfected E. tenella-Ap2 oocysts were expressing e YFP in a nuclear expressing manner. Conclusion: We constructed a transgenic E. tenella line over-expressing EtAp2 successfully. Our results established a basis for the further study on mining the downstream genes regulated by this transcription factor, which might helpful in understanding the mechanism of precocious.
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