[A method for screening microRNA target genes in goat skin by microRNA seed sequence mediated PCR].
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 profile.
Animal Genetics Breeding and Reproduction Autonomous Region Key Laboratories, Inner Mongolia Agricultural University, Hohhot 010018, China. fushao1234@126.com
Shao-Yin
SY
Fu
De-Chao
DC
Zhao
Hong-Li
HL
Zhao
Jin-Quan
JQ
Li
Wen-Guang
WG
Zhang
D000818:Animals; D005260:Female; D006041:Goats / Q000235:genetics*; D035683:MicroRNAs / Q000235:genetics*; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012867:Skin / Q000378:metabolism*; D059467:Transcriptome
D035683:MicroRNAs