利用种子序列检测山羊皮肤中microRNA靶基因分子方法的建立

doi:10.3724/SP.J.1005.2012.00919

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HEREDITAS ›› 2012, Vol. 34 ›› Issue (7): 919-926.doi: 10.3724/SP.J.1005.2012.00919

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A method for screening microRNA target genes in goat skin by microRNA seed sequence mediated PCR

FU Shao-Yin^1,2,3, ZHAO De-Chao^1,2, ZHAO Hong-Li², LI Jin-Quan^1,2, ZHANG Wen-Guang^1,2,3

1. Animal Genetics Breeding and Reproduction Autonomous Region Key Laboratories, Inner Mongolia Agricultural University, Hohhot 010018, China 2. Animal Science College, Inner Mongolia Agricultural University, Hohhot 010018, China 3. Institute of ATCG, Nei Mongol Bio-Information, Hohhot 010020, China

Received:2011-12-21 Revised:2012-02-08 Online:2012-07-20 Published:2012-07-25

Abstract

Abstract: The purpose of the present study was to establish a new microRNA seed mediated controllable genetic operation, namely MicroRNA Targets Finger Print (MTFP), for screening microRNA targets and detecting target gene expression profiles. Based on combining the complementary sequence of seed sequence, upstream and downstream anchor sequence including special adaptor, microRNA targets were amplified by means of the reverse transcription and special two step PCR. The polyacrylamide gel electrophoresis was used to analyze the sizes of amplified microRNA targets and their abundance of expression, which were used to screen specifically expressed genes in different physiological or experimental conditions. Specific target genes were obtained through isolation of DNA fragments and sequencing methods. As an example, by screening the targets of miR-203 in goat skin, five genes were amplified and sequenced with the sizes of 718 bp (JN709494), 349 bp (JN709495), 243 bp (JN709496), 159 bp (JN709497), and 97 bp (JN709498) from goat skin collections. The novel universal MTFP method could be applied for finding microRNA regulation targets or assessing target gene expression pro-file.

Key words: microRNA, expression abundance, microRNA targets finger print (MTFP), goat, skin

FU Shao-Yin, ZHAO De-Chao, ZHAO Hong-Li, LI Jin-Quan, ZHANG Wen-Guang. A method for screening microRNA target genes in goat skin by microRNA seed sequence mediated PCR[J]. HEREDITAS, 2012, 34(7): 919-926.

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A method for screening microRNA target genes in goat skin by microRNA seed sequence mediated PCR

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