Evidence showed that N6-methyladenosine(m^(6)A)modification plays a pivotal role in influencing RNA fate and is strongly associated with cell growth and developmental processes in many species.However,no information r...Evidence showed that N6-methyladenosine(m^(6)A)modification plays a pivotal role in influencing RNA fate and is strongly associated with cell growth and developmental processes in many species.However,no information regarding m^(6)A modification in Eimeria tenella is currently available.In the present study,we surveyed the transcriptome-wide prevalence of m^(6)A in sporulated oocysts and unsporulated oocysts of E.tenella.Methylated RNA immunoprecipitation sequencing(MeRIP-seq)analysis showed that m^(6)A modification was most abundant in the coding sequences,followed by stop codon.There were 3,903 hypermethylated and 3,178 hypomethylated mRNAs in sporulated oocysts compared with unsporulated oocysts.Further joint analysis suggested that m^(6)A modification of the majority of genes was positively correlated with mRNA expression.The mRNA relative expression and m^(6)A level of the selected genes were confirmed by quantitative reverse transcription PCR(RT-qPCR)and MeRIP-qPCR.GO and KEGG analysis indicated that differentially m^(6)A methylated genes(DMMGs)with significant differences in mRNA expression were closely related to processes such as regulation of gene expression,epigenetic,microtubule,autophagy-other and TOR signaling.Moreover,a total of 96 DMMGs without significant differences in mRNA expression showed significant differences at protein level.GO and pathway enrichment analysis of the 96 genes showed that RNA methylation may be involved in cell biosynthesis and metabolism of E.tenella.We firstly present a map of RNA m^(6)A modification in E.tenella,which provides significant insights into developmental biology of E.tenella.展开更多
鸡球虫病是一种由艾美耳属原生动物寄生虫引起的肠道疾病,是影响家禽业的最重要疾病之一。目前,球虫病的控制主要依赖于抗球虫药物的使用,但由于药物的长期广泛使用导致了球虫的严重耐药性。为研究其耐药性机制,本实验室前期对柔嫩艾美...鸡球虫病是一种由艾美耳属原生动物寄生虫引起的肠道疾病,是影响家禽业的最重要疾病之一。目前,球虫病的控制主要依赖于抗球虫药物的使用,但由于药物的长期广泛使用导致了球虫的严重耐药性。为研究其耐药性机制,本实验室前期对柔嫩艾美耳球虫敏感株、地克珠利耐药株、马杜拉霉素耐药株、盐霉素耐药株进行了转录组测序分析,发现与敏感株相比,柔嫩艾美耳球虫微管蛋白β链(Et TBC)仅在地克珠利耐药株高表达。本研究在此基础上,克隆、表达了EtTBC,并初步分析了其功能及与地克珠利的关系;以柔嫩艾美耳球虫敏感株cDNA为模板成功克隆了EtTBC基因,构建了原核表达重组质粒pET28a-Et TBC,并成功诱导表达了重组蛋白r Et TBC;利用qPCR对柔嫩艾美耳球虫敏感株不同发育阶段以及不同虫株(耐药株、敏感株和田间耐药株)的mRNA转录水平进行了分析,结果显示,该基因的转录水平在敏感株未孢子化卵囊阶段最高,且其mRNA转录水平仅在地克珠利耐药株上调;间接免疫荧光定位结果表明该蛋白主要分布在子孢子除折光体外的细胞质和表面,第二代裂殖子的细胞质和表面。展开更多
基金supported by the National Natural Science Foundation of China(31902298)the Shanxi Provincial Key Research and Development Program,China(2022ZDYF126)+2 种基金the Fund for Shanxi“1331 Project”,China(20211331-13)the Science and Technology Innovation Program of Shanxi Agricultural University,China(2017YJ10)the Special Research Fund of Shanxi Agricultural University for High-level Talents,China(2021XG001)。
文摘Evidence showed that N6-methyladenosine(m^(6)A)modification plays a pivotal role in influencing RNA fate and is strongly associated with cell growth and developmental processes in many species.However,no information regarding m^(6)A modification in Eimeria tenella is currently available.In the present study,we surveyed the transcriptome-wide prevalence of m^(6)A in sporulated oocysts and unsporulated oocysts of E.tenella.Methylated RNA immunoprecipitation sequencing(MeRIP-seq)analysis showed that m^(6)A modification was most abundant in the coding sequences,followed by stop codon.There were 3,903 hypermethylated and 3,178 hypomethylated mRNAs in sporulated oocysts compared with unsporulated oocysts.Further joint analysis suggested that m^(6)A modification of the majority of genes was positively correlated with mRNA expression.The mRNA relative expression and m^(6)A level of the selected genes were confirmed by quantitative reverse transcription PCR(RT-qPCR)and MeRIP-qPCR.GO and KEGG analysis indicated that differentially m^(6)A methylated genes(DMMGs)with significant differences in mRNA expression were closely related to processes such as regulation of gene expression,epigenetic,microtubule,autophagy-other and TOR signaling.Moreover,a total of 96 DMMGs without significant differences in mRNA expression showed significant differences at protein level.GO and pathway enrichment analysis of the 96 genes showed that RNA methylation may be involved in cell biosynthesis and metabolism of E.tenella.We firstly present a map of RNA m^(6)A modification in E.tenella,which provides significant insights into developmental biology of E.tenella.
文摘鸡球虫病是一种由艾美耳属原生动物寄生虫引起的肠道疾病,是影响家禽业的最重要疾病之一。目前,球虫病的控制主要依赖于抗球虫药物的使用,但由于药物的长期广泛使用导致了球虫的严重耐药性。为研究其耐药性机制,本实验室前期对柔嫩艾美耳球虫敏感株、地克珠利耐药株、马杜拉霉素耐药株、盐霉素耐药株进行了转录组测序分析,发现与敏感株相比,柔嫩艾美耳球虫微管蛋白β链(Et TBC)仅在地克珠利耐药株高表达。本研究在此基础上,克隆、表达了EtTBC,并初步分析了其功能及与地克珠利的关系;以柔嫩艾美耳球虫敏感株cDNA为模板成功克隆了EtTBC基因,构建了原核表达重组质粒pET28a-Et TBC,并成功诱导表达了重组蛋白r Et TBC;利用qPCR对柔嫩艾美耳球虫敏感株不同发育阶段以及不同虫株(耐药株、敏感株和田间耐药株)的mRNA转录水平进行了分析,结果显示,该基因的转录水平在敏感株未孢子化卵囊阶段最高,且其mRNA转录水平仅在地克珠利耐药株上调;间接免疫荧光定位结果表明该蛋白主要分布在子孢子除折光体外的细胞质和表面,第二代裂殖子的细胞质和表面。
