[Objective] To analyze Eimeria acervulina cSZ1 gene sequence and thus to find a candidate antigen for vaccine development. [Method] According to the cSZ1 gene sequence of Eimeria acervulina published in GenBank, a pai...[Objective] To analyze Eimeria acervulina cSZ1 gene sequence and thus to find a candidate antigen for vaccine development. [Method] According to the cSZ1 gene sequence of Eimeria acervulina published in GenBank, a pair of specific primers was designed. Then, the cSZ1 gene of Eimeria acervulina Changchun strain was amplified by reverse-transcription polymerase chain reaction (RT-PCR) and sequenced. [Result] Compared with the published cSZ1 gene sequence, five nucleotide mutation sites appeared in the cSZ1 gene of Eimeria acervulina Changchun strain. They had a nucleotide sequence similarity of 99.47%, and their encoding region had a nucleotide sequence similarity of 99.61%. Among these mutation sites, two appeared in cSZ1 gene ORF, and A185G mutation led to substitution of valine by isoleucine. [Conclusion] Protein encoded by cSZ1 gene can be used as a candidate antigen for vaccine development.展开更多
为了研究堆型艾美耳球虫(Eimeria acervulina)裂殖子的外分泌蛋白对马-达氏牛肾(MDBK)细胞凋亡的抑制作用,本试验建立Transwell小室间接共培养体系,即将E.acervulina裂殖子与MDBK细胞共培养,采用流式细胞术分别检测共培养1.5、3和6 h M...为了研究堆型艾美耳球虫(Eimeria acervulina)裂殖子的外分泌蛋白对马-达氏牛肾(MDBK)细胞凋亡的抑制作用,本试验建立Transwell小室间接共培养体系,即将E.acervulina裂殖子与MDBK细胞共培养,采用流式细胞术分别检测共培养1.5、3和6 h MDBK细胞的凋亡情况,利用液相色谱串联质谱(LC-MS/MS)对共培养6 h后的细胞上清液中的成分进行鉴定。结果显示,1.5、3和6 h共培养组MDBK细胞的总细胞凋亡率分别为(5.92±0.19)%、(8.13±0.32)%和(11.57±0.37)%,与对照组相比,总细胞凋亡率均极显著降低(P<0.01)。LC-MS/MS分析共鉴定到65个E.acervulina的外分泌蛋白,包括热休克蛋白、微线蛋白、能量代谢酶分子、蛋白磷酸酶分子和未知功能蛋白等。结果表明,E.acervulina裂殖子外分泌蛋白可抑制MDBK细胞凋亡,具体哪种蛋白在抑制细胞凋亡中发挥作用有待后续进一步研究。展开更多
基金funded by the Science and Technology Development Planning Program of Jilin Province (20090578)
文摘[Objective] To analyze Eimeria acervulina cSZ1 gene sequence and thus to find a candidate antigen for vaccine development. [Method] According to the cSZ1 gene sequence of Eimeria acervulina published in GenBank, a pair of specific primers was designed. Then, the cSZ1 gene of Eimeria acervulina Changchun strain was amplified by reverse-transcription polymerase chain reaction (RT-PCR) and sequenced. [Result] Compared with the published cSZ1 gene sequence, five nucleotide mutation sites appeared in the cSZ1 gene of Eimeria acervulina Changchun strain. They had a nucleotide sequence similarity of 99.47%, and their encoding region had a nucleotide sequence similarity of 99.61%. Among these mutation sites, two appeared in cSZ1 gene ORF, and A185G mutation led to substitution of valine by isoleucine. [Conclusion] Protein encoded by cSZ1 gene can be used as a candidate antigen for vaccine development.
