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《Scientia Agricultura Sinica》 2013-06
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Upregulation and Expression of Bombyx mori bmo-miR-14 and Prediction of Its Target Genes

LIU Yue1,YANG Lan-cui2,NIE Zuo-ming2,LU Xuan1,Lü Zheng-bing2,CHEN Jian2,YU Wei2,WU Xiang-fu2,ZHANG Yao-zhou2(1Departmen of Applied Engineering,Zhejiang Economic & Trade Polytechnic,Hangzhou 310018;2Institute of Biochemistry,School of Life Science,Zhejiang Sci-Tech University,Hangzhou 310018)  
【Objective】The objective of this study is to offer a method for upregulating miRNA and predicting its targets in BmN cells,and to provide a methodological reference for studying the function of miRNAs in BmN cells.【Method】An eukaryotic expression vector pSK-hr3-ie1-EGFP-pri-mir-14 for expressing Bombyx mori bmo-miR-14 was constructed and the miRNA gene was expressed in BmN cells.Additionally,the level of bmo-miR-14 in BmN cells was also up-regulated by transfecting chemically synthesized miRNA mimics.Firstly,the fragment containing pri-mir-14 was amplified by PCR and inserted into the downstream of EGFP of eukaryotic expression vector pSK-hr3-ie1-EGFP.The recombinant plasmid pSK-hr3-ie1-EGFP-pri-mir-14,control plasmid pSK-hr3-ie1-EGFP,bmo-miR-14 mimics and negative control mimics were transfected into BmN cells,respectively.The fluorescent light was observed and used to identify the efficiency of transfection.The miRNA levels in transfected BmN cells were identified by qRT-PCR.The RNAhybrid and miRanda were used to predict the targets of bmo-miR-14,respectively.【Result】The miR-14 level was improved in BmN cells by transfecting with pSK-hr3-ie1-EGFP-pri-mir-14 and bmo-miR-14 mimics,and the level was improved as 2.1 and 984 times as the control,respectively.Further,the targeted genes of bmo-miR-14 were identified by bioinformatics tools and a total of 153 and 171 potential targeted genes were identified by RNAhybrid and miRanda software,respectively.In the predicted target genes,a total of 49 genes were predicted by RNAhybrid and miRanda together.The 49 targeted genes were assigned to GO terms of molecular function ontology,and binding and catalytic were overrepresented.In the biological process ontology,cellular process and metabolic process overrepresented among the 49 genes.【Conclusion】The approach to upregulate the level of bmo-miR-14 by transfecting with recombinant expression vector or miRNA mimics has laid a good foundation for the further studies on bmo-miR-14 in BmN cells.
【Fund】: 浙江省自然科学基金项目(LY12C06003 Y3090304);; 国家高技术研究发展计划“863”项目(2011AA100603)
【CateGory Index】: S881.26
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【Citations】
Chinese Journal Full-text Database 2 Hits
1 Wu Jiachang1,Ma Qiong1,Zou Xianfei2,Yang Tongtao1,Shan Lequn1,Ma Baoan1 (1 Orthopaedic Oncology Institute,Tangdu Hospital,the Fourth Military Medical University,Xi’an,Shaanxi,710038 China;2 Ambulance corps of 75135 Unit,Guigang,Guangxi,510710 China);Advance on MicroRNA and Cell Differentiation[J];Progress in Modern Biomedicine;2010-03
2 CHEN Qing-yun,WANG Qing-qing(Institute of Immunology,Collge of Medicine,Zhejiang University,Hangzhou 310058,China);Function of microRNAs in development of immune system and in regulation of immune response[J];Journal of Zhejiang University(Medical Sciences);2010-03
【Co-citations】
Chinese Journal Full-text Database 2 Hits
1 YUAN Zhi-jian 1 ,ZHOU Hong 1 ,HE Xiao-sheng 2 ,HE Wen-juan 1 (1. Wuxi Higher Health Vocational Technical School,Wuxi 214028,China; 2. School of Clinical Medicine,Hangzhou Normal University,Hangzhou 310016,Chian);Differentiation and miRNA expressiom of adipose-derived stem cells in fat transplantation[J];Chinese Journal of Biochemical Pharmaceutics;2012-06
2 Zhang Zi-qiang,Liu Yu-mei,Deng Wen,Lü Qiong-xia College of Animal Science and Technology,Henan University of Science and Technology,Luoyang 471003,Henan Province,China;miRNA and biological behavior of stem cells[J];Journal of Clinical Rehabilitative Tissue Engineering Research;2011-10
【Secondary Citations】
Chinese Journal Full-text Database 2 Hits
1 Li Ming-heng1,2, Zhang Yong2, Li Xiao-yu1, Tian Wei-dong1, Liu Lei1,21State Key Laboratory of Oral Diseases; 2Department of Oral & Maxillofacial Surgery, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, Sichuan Province, China;Changes in microRNA-16 expression during adipogenic differentiation of adipose tissue-derived stromal cells[J];Journal of Clinical Rehabilitative Tissue Engineering Research;2009-02
2 AN Yang,AN Tie-Zhu,TENG Chun-Bo Laboratory of Animal Development Biology,College of Life Science,Northeast Forestry University,Harbin 150040,China;miRNAs play an essential role in stem cell self-renewal and differentiation[J];Hereditas;2009-02
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