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鲤疱疹病毒3型感染的KCF-1细胞中细胞和病毒微小RNA的鉴定与表达分析

Identification and expression analysis of cellular and viral microRNAs in CyHV3-infected KCF-1 cells.

作者信息

Lee Xuezhu, Weng Shaoping, Dong Gang, Zhang Hetong, Zeng Jie, He Jianguo, Dong Chuanfu

机构信息

MOE Key Laboratory of Aquatic Food Safety/State Key Laboratory for Bio-control, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, People's Republic of China; Institute of Aquatic Economic Animals and Guangdong Province Key Laboratory for Aquatic Economic Animals, Sun Yat-sen University, Guangzhou 510275, People's Republic of China.

MOE Key Laboratory of Aquatic Food Safety/State Key Laboratory for Bio-control, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, People's Republic of China; Institute of Aquatic Economic Animals and Guangdong Province Key Laboratory for Aquatic Economic Animals, Sun Yat-sen University, Guangzhou 510275, People's Republic of China; Guangdong Provincial Key Laboratory of Marine Resources and Coastal Engineering, School of Marine Sciences, Sun Yat-sen University, 135 Xingang Road West, Guangzhou 510275, People's Republic of China.

出版信息

Gene. 2016 Oct 30;592(1):154-163. doi: 10.1016/j.gene.2016.07.064. Epub 2016 Jul 29.

Abstract

MicroRNAs (miRNAs) are small non-coding RNAs with approximately 22 nucleotides (nt) that are encoded by a diverse range of metazoan eukaryotes, plants and viruses. CyHV-3 (cyprinid herpesvirus-3) is a member of the Alloherpesviridae virus family and has caused severe economic losses for the common carp and koi carp fishery industries. In this study, a total of 15,987,652 clean reads were generated from a cDNA library of CyHV-3-infected KCF-1 (koi caudal fin) cells using high-throughput sequencing technology. Following annotation and secondary structure prediction, 28 miRNAs were identified as novel candidate miRNAs encoded by common carp (Cyprinus carpio), and seven miRNAs were shown to be encoded by CyHV-3. Next, 19 host miRNAs and seven viral miRNAs were validated by stem-loop real-time PCR. Northern blot analysis confirmed the presence of 14 host miRNAs and five CyHV-3-encoded novel miRNAs. The results of this study expand the knowledge of common carp and CyHV-3 microRNAs and provide a useful theoretical foundation for further study of CyHV-3.

摘要

微小RNA(miRNA)是一类由多种后生动物真核生物、植物和病毒编码的约22个核苷酸(nt)的小非编码RNA。鲤疱疹病毒3型(CyHV - 3)是异疱疹病毒科病毒家族的成员,已给鲤鱼和锦鲤渔业造成了严重的经济损失。在本研究中,使用高通量测序技术从CyHV - 3感染的KCF - 1(锦鲤尾鳍)细胞的cDNA文库中总共生成了15,987,652条干净 reads。经过注释和二级结构预测,鉴定出28个miRNA为鲤鱼(Cyprinus carpio)编码的新型候选miRNA,并且有7个miRNA显示为由CyHV - 3编码。接下来,通过茎环实时PCR验证了19个宿主miRNA和7个病毒miRNA。Northern印迹分析证实了14个宿主miRNA和5个CyHV - 3编码的新型miRNA的存在。本研究结果扩展了对鲤鱼和CyHV - 3微小RNA的认识,并为进一步研究CyHV - 3提供了有用的理论基础。

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