摘要
[目的]鉴定鸡circ_DNAJC3并预测其功能,为进一步研究其在细胞生物学过程中的作用奠定基础.[方法]通过PCR扩增、琼脂糖凝胶电泳、Sanger测序、亚细胞定位、RT-qPCR和生物信息学软件鉴定鸡circ_DNAJC3的环状结构及其稳定性、解析其在细胞中的位置和在不同组织中的表达水平、并预测其功能和可能的作用方式.[结果]鸡circ_DNAJC3由外显子2至4构成稳定的共价环状结构,共311 bp;具有物种保守性;在细胞质中显著性富集(P<0.05);在回肠、胸腺、脾脏、盲肠中高表达(P<0.05)而在下丘脑和小脑中低表达(P<0.05);其可能互作的RNA结合蛋白为AGO2、CELF2、PTBP1和HNRNPC;可能与gga-miR-132b-5p发生互作,其靶基因可能通过钙离子信号通路参与细胞增殖、分化、凋亡及代谢.[结论]揭示了鸡circ_DNAJC3真实存在,可能与RNA结合蛋白和gga-miR-132b-5p互作参与调节细胞生物学过程.
Abstract
[Objective]This study aimed to identify chicken circ_DNAJC3 and predict its potential func-tion in order to lay the foundation for further research on its role in cell biological process.[Method]PCR amplification,agarose gel electrophoresis,Sanger sequencing,subcellular localization,RT-qPCR and bioinformatics software were used to identify the circular structure and stability of circ_DNAJC3,to analyze its location in cells,and its expression level in different tissues,and to predict its potential func-tion and possible modes of action.[Result]Chicken circ_DNAJC3 consists of a stable covalent circular structure from Exons 2 to 4,which is 311 bp.It is conserved in different species,significantly enriched in the cytoplasm(P<0.05),highly expressed in the ileum,thymus,spleen,and cecum(P<0.05)while lower expressed in the hypothalamus and cerebellum(P<0.05).The RNA-binding proteins with which circ_DNAJC3 may interact are AGO2,CELF2,PTBP1 and HNRNPC.Moreover,circ_DNAJC3 can po-tentially interact with gga-miR-132b-5p,and its target genes may participate in cell proliferation,dif-ferentiation,apoptosis,and metabolism through the calcium ion signaling pathway.[Conclusion]These results suggest that chicken circ_DNAJC3 truly exists and may interact with RNA-binding proteins and gga-miR-132b-5p to regulate cellular biological processes.
基金项目
扬州市政策引导类计划(YZ2023260)
江苏省高校"青蓝工程"资助项目()
扬州市第四期英才培育计划优秀教育人才资助项目()
扬州市职业大学校级自然科学项目(2018ZR19)
国家科技期刊平台
NSTL
维普
万方数据