Computational identification and characterization of microRNAs and their targets in Penaeus monodon
Computational identification and characterization of microRNAs and their targets in Penaeus monodon作者机构:Ministry of Education Key Laboratory of Marine Genetics and Breeding College of Marine Life Sciences Ocean University of China Institute of Evolution and Marine Biodiversity Ocean University of China
出 版 物:《Journal of Oceanology and Limnology》 (海洋湖沼学报(英文))
年 卷 期:2018年第36卷第3期
页 面:853-869页
核心收录:
学科分类:0710[理学-生物学] 090801[农学-水产养殖] 0908[农学-水产] 07[理学] 0707[理学-海洋科学] 09[农学] 0815[工学-水利工程] 0903[农学-农业资源与环境] 0713[理学-生态学]
基 金:Supported by the National Natural Science Foundation of China(Nos.31172391,31472274) the Fundamental Research Funds for Central Universities(No.201762003) the Scholarship Foundation for Excellent Scientists of Shandong Province(No.BS2011SW054) the National HighTech R&D Program of China(863 Program)(No.2012AA10A402) the Open Funds of Institute of Biodiversity and Evolution,Ocean University of China(No.201362017)
摘 要:Study on shrimp miRNAs was limited and just 7 mature miRNA sequences of Marsupenaeus japonicus are deposited in mir Base database. In this study, miRNAs and their target gene candidates were computationally identified from shrimp Penaeu s monodon and then experimentally validated. Using 39 908 expressed sequence tags(ESTs) and 21 124 genome survey sequences(GSSs) of P. monodon(pmo) as reference dataset, a comprehensive approach based on inter-species homolog search was employed to investigate the candidate miRNAs(i.e. pmo-miRNA). A total of eight miRNAs belonging to 7 families were computationally identified and five out of them were subsequently validated by PCR and sequencing. Of these, pmo-miR-4961a, pmo-miR-4961b, pmo-miR-4979 and pmo-miR-3819 were first identified from shrimps. Both the mature pmo-miRNAs and the corresponding precursors were conserved among different species. Based on perfect or near-perfect match to the target region, the target gene candidates of pmomiRNAs were predicted from 10 331 mRNA sequences of P. monodon. A total of 20 genes were predicted as the targets of pmo-miR-4961a, pmo-miR-4961b, pmo-miR-4979 and pmo-miR-6492. Experimental validation by dual luciferase reporter assay confi rmed the targeting between 3 pmo-miRNAs and one or two of their target genes, especially the pmo-miR-4979 which could significantly down-regulate the expression of target gene(JR226772). This study updates the miRNAs and their targets in P. monodon and lays a solid foundation for future RNAi study.