数字农科院2.0

弓形虫HAD2b基因缺失株的构建及生物学功能研究

文献类型： 中文期刊

作者： 张海生;梁勤立;李婷婷;朱兴全;王金磊

作者机构：

关键词： 弓形虫;RHΔHAD2b;CRISPR-Cas9;生物学功能

期刊名称：中国兽医科学

ISSN： 1673-4696

年卷期： 2021 年 51.0 卷 5.0 期

页码：

收录情况： 北大核心（2020版） ; CSCD（2020-2021年度） ; 农林核心（2020版） ; 科技核心（2020版）

摘要： 弓形虫是一种很成功的专性细胞内寄生的机会性致病原虫,可感染几乎所有温血动物,导致的弓形虫病是一种重要的人兽共患寄生虫病,目前尚无有效的疫苗。为了研究弓形虫卤代酸脱卤酶(haloacid dehalogenase,HAD)家族中一个保守蛋白HAD2b的功能,本研究通过利用CRISPR/Cas9技术成功构建了弓形虫Ⅰ型RH虫株HAD2b基因缺失株(RHΔHAD2b)。通过噬斑试验分析发现,RHΔHAD2b形成的噬斑大小和数量与野生株相比无显著差异。通过逸出试验发现,大部分RHΔHAD2b和野生株在钙离子的刺激下,在2 min内完成逸出,差异不显著。小鼠毒力试验发现,昆明鼠分别感染RHΔHAD2b和野生株后均在7～10 d内死亡,无显著差异。对HAD2b基因转录组数据库现有数据分析表明,HAD2b在弓形虫RH株的表达量显著低于其它虫株,说明HAD2b在不同虫株可能发挥不同的功能。本研究成功构建了弓形虫HAD2b基因缺失株(RHΔHAD2b),为进一步研究HAD2b基因在Ⅰ型虫株的其它生物学功能奠定了基础,也为构建弓形虫其它基因型虫株的HAD2b基因缺失株奠定了技术基础。

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