靶向猪ATGL 基因的miRNA预测及鉴定

doi:10.11843/j.issn.0366-6964.2015.08.002

Abstract

Abstract:

To preliminary screen out ssc-miRNAs targeting porcine adipose triglyceride lipase gene (ATGL)，10 ssc-miRNAs targeting ATGL were predicted by bioinformatics analysis．3′UTR of porcine ATGL gene was inserted into the pMIR-Luc target reporter vector and transiently co-transfected with candidate miRNA mimics into HEK 293T cells，using pRL-TK vector as an internal control reporter.Three other reporter vectors containing the mutated ssc-miRNAs targeting sites by reversing the seed region of targeting sites were created as negative controls.The dual luciferase reporter assay system was used to evaluate the activity of luciferase．The results showed that pMIR-ATGL-3′UTR recombinant vector was successfully constructed which contained 3′UTR of porcine ATGL gene.Dual luciferase reporter assay showed that ssc-miR-769-3p，ssc-miR-1343，ssc-miR-7137-3p，ssc-miR-671-5p and ssc-miR-149 could down-regulate the luciferase activity of pMIR-ATGL-3′UTR vector in 293T cell，while mutant assay verified ssc-miR-1343 and ssc-miR-7137-3p could down-regulate the luciferase activity by targeting the predicted seed region of 3′UTR of porcine ATGL gene.These results indicated that ssc-miR-1343 and ssc-miR-7137-3p could down-regulate the expression of porcine ATGL gene by targeting its 3′UTR.

CLC Number:

S828.2

DAI Li-he，CHU Xiao-hong，LU Fu-zeng，HUANG Sun-ping，XU Ru-hai. Prediction and Validation of miRNA Targeting Porcine ATGL Gene[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(8): 1281-1289.

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本刊相关报道（以下文章由本刊网站www.xmsyxb.com可以免费下载查阅）：
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Prediction and Validation of miRNA Targeting Porcine ATGL Gene

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