RNA干扰abcg-5基因对犬弓首蛔虫感染性的影响

doi:10.11843/j.issn.0366-6964.2019.02.017

畜牧兽医学报 ›› 2019, Vol. 50 ›› Issue (2): 390-397.doi: 10.11843/j.issn.0366-6964.2019.02.017

RNA干扰abcg-5基因对犬弓首蛔虫感染性的影响

罗永莉^1,2, 江艾耘¹, 胡志刚¹, 李相兰¹, 杨晓迪¹, 旷策嫣¹, 周荣琼^1*

1. 西南大学动物科学学院, 荣昌 402460;
2. 重庆三峡职业学院, 万州 404155

收稿日期:2018-06-29 出版日期:2019-02-23 发布日期:2019-02-23
通讯作者: 周荣琼,主要从事畜禽寄生虫病学及分子寄生虫学研究,E-mail:rongqiongzhou@126.com
作者简介:罗永莉(1992-),女,重庆人,硕士生,主要从事寄生虫分子生物学的研究,E-mail:luoyongli21@163.com
基金资助:
国家自然科学基金（31672541；31172313）；中央高校基本科研业务（XDJK2016E087；XDJK2017D080）

Effect of RNA Interference abcg-5 Gene on the Infectivity of Toxocara canis

LUO Yongli^1,2, JIANG Aiyun¹, HU Zhigang¹, LI Xianglan¹, YANG Xiaodi¹, KUANG Ceyan¹, ZHOU Rongqiong^1*

1. College of Animal Science, Southwest University, Rongchang 402460, China;
2. Chongqing Three Gorges Vocational College, Wanzhou 404155, China

Received:2018-06-29 Online:2019-02-23 Published:2019-02-23

摘要/Abstract

摘要：

为研究犬弓首蛔虫（Toxocara canis，T.canis）腺苷三磷酸结合盒G5基因（ATP-binding cassette G5，abcg-5）的功能，通过体外合成Tc-abcg-5基因的特异性siRNA，利用RNAi技术对T.canis的成虫和虫卵进行干扰。结果显示，干扰24、48 h后siRNA-744组Tc-abcg-5的相对表达量比siRNA-1020组和siRNA-432组显著性降低，表明siRNA-744有较高的沉默效率。采用电穿孔法和浸泡法对siRNA-744的递送效率进行比较，结果显示浸泡法更有利于将siRNA-744导入到T.canis虫卵内。用siRNA-744经浸泡法干扰T.canis感染性虫卵，经口灌服小鼠，感染后第7天对小鼠的脑、肝和肺组织进行病理学研究，结果显示RNA干扰使T.canis感染性虫卵的感染力下降，从而降低了肺、肝的病变程度。研究表明Tc-abcg-5基因可能通过影响虫卵的发育从而参与T.canis的生长、发育过程。

Abstract:

This experiment was conducted to study the effect of ATP-binding cassette G5 gene of Toxocara canis. Specific siRNAs of the Tc-abcg-5 gene fragment were synthesized in vitro, and adult T. canis and eggs were interfered by using RNAi techniques. Results were as follows:Compared with siRNA-1020 group and siRNA-432 group, the transcriptional level of Tc-abcg-5 in siRNA-744 group was significantly reduced after 24 and 48 h's treatments, indicating that siRNA-744 had higher silencing efficiency. The delivery efficiency of siRNA-744 to eggs was compared by electroporation and immersion methods. The results showed that immersion method was more efficient. Interference of T. canis infective eggs were performed using siRNA-744 by soaking. Pathology study was used to analysis of mouse liver, brain and lung tissues on the 7th day after oral administration of interfered eggs, which showed that RNAi reduced the infectivity of T. canis infective eggs, thereby reducing the lung and liver lesions. It is suggested that the Tc-abcg-5 gene may participate in the growth and development of T. canis by affecting the development of eggs.

中图分类号:

S852.731

罗永莉, 江艾耘, 胡志刚, 李相兰, 杨晓迪, 旷策嫣, 周荣琼. RNA干扰abcg-5基因对犬弓首蛔虫感染性的影响[J]. 畜牧兽医学报, 2019, 50(2): 390-397.

LUO Yongli, JIANG Aiyun, HU Zhigang, LI Xianglan, YANG Xiaodi, KUANG Ceyan, ZHOU Rongqiong. Effect of RNA Interference abcg-5 Gene on the Infectivity of Toxocara canis[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(2): 390-397.

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RNA干扰abcg-5基因对犬弓首蛔虫感染性的影响

Effect of RNA Interference abcg-5 Gene on the Infectivity of Toxocara canis

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