[1]赵园园,孟金柱.miR-27a与SCF 靶向关系鉴定[J].江苏农业学报,2018,(02):271-275.[doi:doi:10.3969/j.issn.1000-4440.2018.02.006]
　ZHAO Yuan-yuan,MENG Jin-zhu.Identification of targeted relationship between miR-27a and SCF[J].,2018,(02):271-275.[doi:doi:10.3969/j.issn.1000-4440.2018.02.006]
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miR-27a与SCF 靶向关系鉴定()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2018年02期
页码:
271-275
栏目:
遗传育种·生理生化
出版日期:
2018-04-25

文章信息/Info

Title:
Identification of targeted relationship between miR-27a and SCF
作者:
赵园园; 孟金柱
(铜仁学院乌江学院，贵州铜仁554300)
Author(s):
ZHAO Yuan-yuan; MENG Jin-zhu
(Wujiang College of Tongren University, Tongren 554300, China)
关键词:
miR-27a; SCF; 靶向关系
Keywords:
miR-27a; SCF; targeted relationship
分类号:
Q786
DOI:
doi:10.3969/j.issn.1000-4440.2018.02.006
文献标志码:
A
摘要:
为确定miR-27a与SCF的靶向关系，剪取成年棕色小鼠和灰色小鼠的皮肤组织，qRT-PCR 检测 miR-27a 和 SCF 的表达量,选用 PicTar、TargetScan 和 DIANA-microT 3个数据库对 miR-27a 进行靶基因预测,构建 miR-27a 的真核表达载体和 SCF 3′UTR （含种子序列）的双荧光表达载体，并对种子序列进行突变处理,以 miR-27a 过表达载体或 NC 载体（空载）组合 SCF 3′UTR 双荧光报告载体或 SCF 3′UTR 的突变载体共转染 HEK 293 T 细胞，48 h 后检测双荧光素酶活性。qRT-PCR 结果显示，miR-27a 在灰色小鼠皮肤中的表达量极显著高于棕色小鼠（P<0.01）,而 SCF 在棕色小鼠皮肤中的表达量显著高于灰色小鼠（P<0.05）；3个数据库的分析结果均显示，SCF 是 miR-27a 的靶基因；双荧光报告试验结果显示，miR-27a 能显著抑制双荧光活性，即 miR-27a 能与 SCF 3′UTR 的种子序列结合。综上所述，SCF 是 miR-27a 的靶基因。
Abstract:
To determine the targeted relationship of miR-27a and stem cell factor(SCF), skin tissues were obtained from gray and brown adult mouse and qRT-PCR was performed to detect the expression of miR-27a and SCF. PicTar, TargetScan and DIANA-microT databases were chosen to predict the targeted genes of miR-27a. Eukaryotic expression vector of miR-27a and double fluorescent report vector of SCF 3′UTR containing seed sequence were constructed, respectively. Mutation was performed for the seed sequence of 3′UTR of SCF. The miR-27a over expression vector or NC vector (no-load) combined double fluorescent report vector of SCF 3 ′UTR or mutated vector of SCF 3′UTR was co-transfected into HEK 293 T cells, and dual luciferase activity was detected after 48 hours. The results of qRT-PCR showed that the expression level of miR-27a in gray mice skin were extremely significantly greater than that in brown mice (P<0.01). Whereas, the expression level of SCF mRNA in brown mice skin was significantly higher than that in gray mice (P<0.05). Bioinformatics analysis of the three databases showed that SCF was a targeted gene of miR-27a. Double fluorescent report test results showed that miR-27a could significantly inhibit the double fluorescence activity. Namely，miR-27a could combine with seed sequence of SCF 3′UTR. In conclusion, SCF is a targeted gene of miR-27a.

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备注/Memo

备注/Memo:
收稿日期：2017-10-17 基金项目：铜仁学院博士启动基金项目（trxyDH1601）；铜仁市科技局计划项目[铜市科研(2016)18 号-1] 作者简介：赵园园（1987-），女，河北邢台人，博士，副教授，研究方向为动物遗传，（ E-mail）84840293@163.com
更新日期/Last Update: 2018-05-04