文章摘要
常规培养与缺氧条件下BCG菌体内外sRNA的表达检测
sRNA Expression Test in vivo and in vitro of BCG under Routine Culture and Hypoxia
  
DOI:doi:10.3969/j.issn.1005-7021.2021.02.008
中文关键词: 卡介苗(BCG)  细胞外RNA  细菌sRNA  缺氧  RT-qPCR
英文关键词: BCG  in vitro RNA  bacterial sRNA  hypoxia  RT-qPCR
基金项目:国家科技重大专项(2017ZX10201301-003-005)
作者单位
韩雪 哈尔滨医科大学 微生物学教研室黑龙江 哈尔滨 150081 
顾伟 哈尔滨医科大学 微生物学教研室黑龙江 哈尔滨 150081 
李婷 哈尔滨医科大学 微生物学教研室黑龙江 哈尔滨 150081 
范云帆 哈尔滨医科大学 微生物学教研室黑龙江 哈尔滨 150081 
张文莉 哈尔滨医科大学 微生物学教研室黑龙江 哈尔滨 150081 
赵吉子 哈尔滨医科大学 微生物学教研室黑龙江 哈尔滨 150081 
付英梅 哈尔滨医科大学 微生物学教研室黑龙江 哈尔滨 150081 
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中文摘要:
      近期发现细菌的sRNA在菌体内和菌体外均具有一定的生物学功能。为研究结核分枝杆菌菌体内外sRNA的表达情况,通过分析卡介苗(Bacillus Calmette-Guerin Vaccine,BCG)菌体和外泌体RNA测序结果,采用RT-qPCR法检测常规培养与缺氧条件下BCG菌体内外sRNA相对表达量,分析菌体内外sRNA谱的差异。结果显示,常规培养时,菌体内丰度较高的sRNA为MTS2823、MTS1338与ASdes,菌体外丰度较高的为Mcr3、MTS2823和AS1890。菌体内受缺氧诱导表达增加的是MTS2823、MTS1338、MTS0997和G2。其中MTS1338与G2的启动子区发现了DosR的结合基序。无论是否缺氧,MTS0997、G2、Mcr7和AS1890在菌体外的相对表达量均高于菌体内,而C8只在缺氧时菌体外表达增高。研究揭示了BCG菌体内外sRNA表达谱不同,且一部分sRNA在常规培养和缺氧应激时向菌体外释放,同时发现了细菌受缺氧诱导的sRNA种类。
英文摘要:
      Recently it has been found that bacterial sRNA can play a certain biological function both in vivo and in vitro thallus. In order to study the expression of in vivo and in vitro sRNA of Mycobacterium tuberculosis, the results of RNA sequencing of BCG thallus and exosomes were analyzed. The relative expression of in vivo and in vitro sRNA in BCG thallus under routine culture and hypoxia was detected by RT-qPCR method, and the difference of in vivo and in vitro sRNA genealogies was analyzed. The results showed that when culture routinely, the higher thallus in vivo abundance were MTS2823, MTS1338, and ASdes. The higher thallus in vitro abundance was Mcr3, MTS2823, and AS1890. The induced expression increased in vivo by hypoxia was MTS2823, MTS1338, MTS0997, and G2 was induced by hypoxia. Among them the binding motif sequence of DosR was found in the promoter region of MTS1338 and G2. Whether hypoxia or not, the relative expression of MTS0997, G2, Mcr7, and AS1890 in vitro thallus was higher than that in vivo thallus, and the expression of C8 increased when in hypoxia. This study had revealed that in vivo and in vitro sRNA expression genealogies of BCG thallus were different. And part of sRNA was released to in vitro thallus during routine culture and hypoxia stress, and it was found simultaneously that the types of bacterial sRNA induced by hypoxia.
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