摘要
为揭示弓形虫突触融合蛋白STXQa1在虫体生长过程中的作用,本研究利用ddFKBP系统构建了STXQa1的条件性过表达虫株。利用间接免疫荧光试验(IFA)观察STXQa1的亚细胞定位,通过westen blot及IFA检测STXQa1蛋白的表达;经IFA检测STXQa1蛋白过表达虫株在细胞内的增殖能力、侵入细胞的能力以及逸出细胞的能力。结果显示STXQa1定位在虫体空泡样隔室(VAC);利用shield1能够快速调控ddFKBP-STXQa1融合蛋白的表达;过表达STXQa1对弓形虫的细胞内增殖具有抑制作用,同时影响弓形虫的侵入及逸出能力。本研究通过对弓形虫独特的突触融合蛋白STXQa1的表达与鉴定,为进一步研究弓形虫分泌细胞器的成熟和蛋白的分泌机制研究奠定了基础。
In order to reveal the role of STXQa1 on growth of Toxoplasma gondii,A parasite strain conditionally overexpressing STXQa1 was constructed based on a ddFKBP system.The expression of STXQa1 was detected by western blott and IFA.The subcellular localization of STXQa1 was investigated by IFA.The ability of replication,invasion and egress of parasites overexpressing STXQa1 was also detected by IFA.The results indicated that STXQa1 localized at the vacuolar compartment(VAC)of T.gondii.The expression of ddFKBP-STXQa1 fusion protein could be regulated tightly by adding Shield1 in cell culture.Overexpression of STXQa1 is toxic to growth of T.gondii and affects invasion and replication of the parasites.In this study,the unique synaptic fusion protein of T.gondii,STXQa1 was identified.The data laid a foundation for the further study on the maturation of secretory organelles and secretion mechanism of T.gondii.
作者
付佳雯
陈赫明
杨娟
张朝霞
郑君
曹世诺
王彩云
贾洪林
FU Jia-wen;CHEN He-ming;YANG Juan;ZHANG Zhao-xia;ZHENG Jun;CAO Shi-nuo;WANG Cai-yun;JIA Hong-lin(College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot 010018,China;State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150069,China)
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2019年第7期689-695,共7页
Chinese Journal of Preventive Veterinary Medicine
基金
国家自然科学基金资助项目(31702230)
黑龙江省留学回国人员择优资助项目(2017QD0024)
兽医生物技术国家重点实验室开放课题基金项目资助(SKLVBF201808)
