微小隐孢子虫氨基酸转运蛋白CpAAT1的亚细胞定位研究  

SUBCELLULAR LOCALIZATION OF AMINO ACID TRANSPORTER CPAAT1 IN CRYPTOSPORIDIUM PARVUM

作　　者：李敏 张颖 王东强[1] 尹继刚[1] 朱冠 LI Min;ZHANG Ying;WANG Dong-qiang;YIN Ji-gang;ZHU Guan(State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases,Institute of Zoonioses/College of Veterinary Medicine,Jilin niversity,Changchun 130062,Jilin,China)

机构地区：[1]人畜共患传染病重症诊治全国重点实验室,吉林大学人兽共患病研究所/动物医学学院,长春130062

出　　处：《寄生虫与医学昆虫学报》2024年第3期137-144,共8页Acta Parasitologica et Medica Entomologica Sinica

基　　金：“十四五”国家重点研发计划资助(2022YFD1800200)。

摘　　要：目的探究微小隐孢子虫氨基酸转运蛋白的亚细胞定位。方法本研究以微小隐孢子虫氨基酸转运蛋白1(Cryptosporidium parvum Amino acid transporter 1,CpAAT1)为研究对象,设计合成了特异性多肽片段,免疫家兔制备多克隆抗体。利用Western blot验证该抗体的特异性,通过间接免疫荧光的方法对该蛋白的亚细胞定位进行了分析。结果CpAAT1多克隆抗体能特异性识别虫体天然蛋白,后者在微小隐孢子虫生活史的各个时期均有表达。子孢子时期,CpAAT1定位于虫体表膜;无性生殖期时该蛋白定位在成熟的裂殖子表膜;在有性生殖阶段,雌雄配子体均可被该抗体标记。结论本研究为进一步探索微小隐孢子虫氨基酸转运蛋白的转运机制提供了一些新线索。Objectives To investigate the subcellular localization of amino acid transporter 1(CpAAT1)in Cryptosporidium parvum.Methods Based on bioinformatics analysis of the amino acid sequence of CpAAT1,specific polypeptide fragments were designed and synthesized for rabbit polyclonal antibodies.The specificity of the antibody was verified using western blotting and the subcellular localization of the protein was analyzed using an indirect immunofluorescence assay(IFA).Results The CpAAT1 polyclonal antibody specifically recognized the native protein of C.parvum and was expressed in all life stages of the parasite.At the sporozoite stage,CpAAT1 was localized on the surface of sporozoites,and at the meront stage,the protein was localized on the surface of the mature merozoite membrane.At the sexual reproduction stage,both male and female gametes can be labeled with the antibody.Conclusions This study provides novel insights that are useful for further exploration of the mechanisms of amino acid transporters in C.parvum.

关 键 词：微小隐孢子虫 氨基酸转运蛋白 间接免疫荧光 亚细胞定位 

分 类 号：R382.3[医药卫生—医学寄生虫学]