马立克病病毒miR-M4-5p宿主靶基因cDNA文库的构建及鉴定

doi:10.11843/j.issn.0366-6964.2018.02.014

Abstract

Abstract:

Marek's disease virus (MDV) is one of the oncogenic herpes viruses that can induce tumors in their natural hosts, which is considered to be an excellent model for investigating the biology, genetics, and immunology of tumorigenesis. Recently, a lot of microRNAs (miRNAs) have been identified in MDV genomes and the miR-M4-5p encoded by MDV-1 has been identified as a viral analog of cellular miR-155. Since miR-155 is a host miRNA associated with several cancers, miR-M4-5p may play a critical role in MDV oncogenesis. The present work was performed to construct a cDNA library and to primarily screen the putative host mRNA targets for miR-M4-5p. All hybrid-PCR products amplified from CEF RNA were harvested and cloned into pMD19-T vectors, then transformed into E. coli JM109 to produce a pool. The clones were selected and sequenced, using the online basic local alignment search tool (BLAST) to analyze the target sequences. A total of 88 candidate mRNA genes were obtained, 29 of which contained the predicted binding miRNA sites in the 3'-UTRs, complementary to the seed sequence of miR-M4-5p. After three rounds of dual fluorescence reporter assays (DLRA), five host genes including PRICKLE1, COLA, BCAT1, ANTXR1 and TECPR1 were primarily identified as the biological targets for miR-M4-5p. Our work provides an important basis for further studies on the molecular regulatory mechanism mediated by miR-M4-5p.

CLC Number:

S852.659.1

XUE Zheng-fei, TENG Man, LI Hui-zhen, MA Sheng-ming, SONG Li-na, ZHANG Ya, LUO Jun, ZHANG Gai-ping. Construction and Identification of the cDNA Library of Host mRNA Targets Recognized by miR-M4-5p Encoded by Marek's Disease Virus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(2): 348-359.

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Construction and Identification of the cDNA Library of Host mRNA Targets Recognized by miR-M4-5p Encoded by Marek's Disease Virus

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