弓形虫不同发育时期棒状体颈部蛋白5基因的表达研究

doi:10.16431/j.cnki.1671-7236.2017.02.007

《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (2): 350-356.doi: 10.16431/j.cnki.1671-7236.2017.02.007

弓形虫不同发育时期棒状体颈部蛋白5基因的表达研究

何帅^1,2, 周春雪^2,3, 朱兴全², 周东辉², 徐前明¹

1. 安徽农业大学动物科技学院, 合肥 230036;
2. 中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, 兰州 730046;
3. 中国农业大学动物医学院, 国家动物寄生原虫实验室, 北京 100193

修回日期:2016-11-15 出版日期:2017-02-20 发布日期:2017-02-25
通讯作者: 周东辉, 徐前明 E-mail:zhoudonghui@caas.cn;vetqmxu@163.com
作者简介:何帅(1990-), 男, 陕西汉中人, 硕士, 研究方向:兽医寄生虫病学及分子生物学, E-mail:heshuaichn@outlook.com
基金资助:
国家自然科学基金项目（31302085）

Study on Expression of Toxoplasma gondii Rhoptry Neck Protein 5 Gene in Different Developmental Stages

HE Shuai^1,2, ZHOU Chun-xue^2,3, ZHU Xing-quan², ZHOU Dong-hui², XU Qian-ming¹

1. College of Animal Science, Anhui Agricultural University, Hefei 230036, China;
2. State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China;
3. National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China

Revised:2016-11-15 Online:2017-02-20 Published:2017-02-25

摘要/Abstract

摘要：

为探究不同发育时期弓形虫棒状体颈部蛋白5（Toxoplasma gondii rhoptry neck protein 5，TgRON5）基因的表达情况与其毒力的相关性，本试验以Ⅱ型弓形虫PRU虫株作为研究对象，以弓形虫管家基因ACT1作为内参基因，分别对目的基因和内参基因设计特异性引物，通过对各基因建立标准曲线，确定二者扩增效率的一致性后，采用实时荧光定量PCR相对定量法对弓形虫4个不同发育时期（速殖子、缓殖子、未孢子化卵囊及孢子化卵囊）中TgRON5基因的表达情况进行检测与分析。结果显示，TgRON5基因在弓形虫各发育时期均有表达，其中，孢子化卵囊表达水平最高，未孢子化卵囊次之，速殖子较低，缓殖子最低，表明RON5蛋白的合成与分泌与虫体入侵宿主细胞的侵袭力有着密切的关系。本研究结果为阐明TgRON5蛋白参与弓形虫入侵的机理奠定了基础。

关键词: 弓形虫; 棒状体颈部蛋白5; 实时荧光定量PCR; 表达水平

Abstract:

To investigate the relationship between the expression level of Toxoplasma gondii rhoptry neck protein 5 (TgRON5) gene in different developmental stages and the virulence of Toxoplasma gondii, the objective of this study was to examine the different expression of TgRON5 gene in different developmental stages of type Ⅱ Toxoplasma gondii PRU strain. Specific primers were designed according to the sequence of TgRON5 gene, and ACT1 gene of Toxoplasma gondii was used as a reference gene. Following the establishment of standard curve for the target and reference genes, in order to confirm the consistency of their amplification efficiency, Real-time PCR method was applied to determine and compare the expression level of TgRON5 gene in development stage which including tachyzoite, bradyzoite, non-sporulated oocyst and sporulated oocyst. The results demonstrated that TgRON5 was expressed in all developmental stages but the sporulated oocysts had the highest expression, followed by the non-sporulated oocysts, the third was tachyzoite, and the lowest was bradyzoite. It suggests that the synthesis and secretion of TgRON5 protein was closely associated with the invasiveness of the parasite. All these findings had important implications for elucidating the functions of TgRON5 involved in the invasion of Toxoplasma gondii.

Key words: Toxoplasma gondii; rhoptry neck protein 5; Real-time PCR; expression level

中图分类号:

Q786

何帅, 周春雪, 朱兴全, 周东辉, 徐前明. 弓形虫不同发育时期棒状体颈部蛋白5基因的表达研究[J]. 《中国畜牧兽医》, 2017, 44(2): 350-356.

HE Shuai, ZHOU Chun-xue, ZHU Xing-quan, ZHOU Dong-hui, XU Qian-ming. Study on Expression of Toxoplasma gondii Rhoptry Neck Protein 5 Gene in Different Developmental Stages[J]. , 2017, 44(2): 350-356.

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弓形虫不同发育时期棒状体颈部蛋白5基因的表达研究

Study on Expression of Toxoplasma gondii Rhoptry Neck Protein 5 Gene in Different Developmental Stages

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