[1]肖春婷,廉传江,易诚,等.Gga-miR-7b和靶基因SNCA在两品系鸡脾脏中的表达分析[J].江苏农业学报,2019,(04):874-879.[doi:doi:10.3969/j.issn.1000-4440.2019.04.018]
　XIAO Chun ting,LIAN Chuan jiang,YI Cheng,et al.Expression analysis of GgamiR7b and target gene SNCA in spleen of two strains of chicken[J].,2019,(04):874-879.[doi:doi:10.3969/j.issn.1000-4440.2019.04.018]
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Gga-miR-7b和靶基因SNCA在两品系鸡脾脏中的表达分析()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2019年04期
页码:
874-879
栏目:
畜牧兽医·水产养殖
出版日期:
2019-08-31

文章信息/Info

Title:
Expression analysis of GgamiR7b and target gene SNCA in spleen of two strains of chicken
作者:
肖春婷1; 2; 廉传江2; 易诚3; 陈洪岩1; 2
（1.东北农业大学生命科学学院,黑龙江哈尔滨150030；2.中国农业科学院哈尔滨兽医研究所/兽医生物技术国家重点实验室/黑龙江省实验动物与比较医学重点实验室,黑龙江哈尔滨150069；3.东北林业大学生命科学学院,黑龙江哈尔滨150040）
Author(s):
XIAO Chunting1; 2; LIAN Chuanjiang2; YI Cheng3; CHEN Hongyan1; 2
（1.College of Life Science, Northeast Agricultural University, Harbin 150030, China；2.Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences/State Key Laboratory of Veterinary Biotechnology/Heilongjiang Provincial Key Laboratory of Laboratory Animal and Comparative Medicine, Harbin 150069, China；3.College of Life Science, Northeast Forestry University, Harbin 150040, China）
关键词:
ggamiR7b; SNCA; 靶向关系; 表达分析
Keywords:
ggamiR7b; SNCA; targeting relationship; expression analysis
分类号:
Q786
DOI:
doi:10.3969/j.issn.1000-4440.2019.04.018
文献标志码:
A
摘要:
为确定 ggamiR7b 与SNCA的靶向关系以及检测他们在MDV超强毒株Md5感染SPF抗性鸡（B21单倍型）和易感鸡（B19单倍型）后5 d、14 d、25 d脾脏中的表达量。利用TargetScan、miRBD软件预测ggamiR7b的靶基因,通过双荧光素酶报告基因检测系统体外验证ggamiR7b与SNCA之间的靶向关系。qRTPCR检测SNCA和ggamiR7b在B21、B19感染Md5脾脏中的表达量。荧光素酶活性结果显示,ggamiR7b与SNCAwt3′UTR共转染后显著抑制萤火虫荧光素酶活性（P＜005）,对SNCAmut3′UTR活性无显著影响。qRTPCR结果显示,SNCA在B21、B19中,Md5感染5 d均显著上调表达（P＜005）,Md5感染25 d时SNCA在B21中极显著上调表达（P＜001）,在B19中极显著下调表达（P＜001）。ggamiR7b在Md5感染14 d,在B21中显著上调表达而在B19中显著下调表达（P＜005）,Md5感染25 d,ggamiR7b在B21、B19中均极显著上调表达（P＜001）。综上所述,SNCA是ggamiR7b的靶基因。ggamiR7b和SNCA可能与MD抗性/易感性以及MD肿瘤转化有关。
Abstract:
The aim of this study was to determine the targeting relationship between ggamiR7b and SNCA and to detect their expression in the spleen of SPF resistant chickens (B21 haplotype) and susceptible chickens (B19 haplotype) infected with MDV supervirus strain Md5 at 5 d, 14 d, and 25 d after infection. The target gene of ggamiR7b was predicted by TargetScan and miRBD software, and the targeting relationship between ggamiR7b and SNCA was verified in vitro by the dual luciferase reporter gene detection system. qRTPCR was used to detect the expression levels of SNCA and ggamiR7b in the spleen of B21 and B19 infected with Md5. The results of luciferase activity showed that the cotransfection of ggamiR7b and SNCAwt3′UTR significantly inhibited firefly luciferase activity (P＜0.05), but had no significant effect on SNCAmut3′UTR activity. The results of qRTPCR showed that the expression of SNCA was significantly upregulated in B21 and B19 on the 5th day after Md5 infection (P＜0.05), and the expression of SNCA in B21 was highly upregulated on the 25th day after Md5 infection(P＜0.05), and significantly downregulated in B19 (P＜0.01). The expression of ggamiR7b was significantly upregulated in B21 and downregulated in B19 on the 14th day after Md5 infection (P＜0.05), and the expression of ggamiR7b was highly upregulated in B21 and B19 on the 25th day after Md5 infection (P＜0.01). In summary, SNCA is the target gene of ggamiR7b. The ggamiR7b and SNCA may be related to MD resistance/susceptibility and MD tumor transformation.

参考文献/References:

[1]KHEIMAR A,PREVIDELLI R L,WIGHT D J,et al. Role in Mareks disease virus pathogenesis, integration and tumorigenesis[J]. Viruses,2017,9(7):173.
[2]BROWN A C,BAIGENT S J,SMITH L P,et al.Interaction of MEQ protein and Cterminalbinding protein is critical for induction of lymphomas by Marek′s disease virus [J].Proc Natl Acad Sci USA,2006,103(6):1687-1692.
［3]陈蓉,应诗家,陈哲,等. 两品种鸭下丘脑 miRNA 的鉴定及其表达验证分析［J].江苏农业学报,2017,33(5):1093-1098.
[4]李会珍,滕蔓,党露,等. 马立克氏病病毒编码的miRM125p对鸡HVCN1基因表达的靶向调控[J]. 河南农业科学,2016,45(6):121-125,129.
[5]李新. miR26a和miR181a在鸡马立克氏病肿瘤形成过程中的作用机制研究[D].北京：中国农业大学,2014.
[6]LEE H J,PALKOVITS M,YONNG W S. miR7b, a microRNA upregulated in the hypothalamus after chronic hyperosmolar stimulation, inhibits Fos translation[J].Proc Natl Acad Sci USA, 2006, 103(42):15669-15674.
[7]YANG X,GAO W,LIU H,et al. MicroRNA transcriptome analysis in chicken kidneys in response to differing virulent infectious bronchitis virus infections [J]. Archives of Virology, 2017, 162(11): 3397-3405.
[8]易诚. gga-miR-21-5p靶向调控SATB1基因及其对马立克氏病肿瘤细胞增殖与迁移的影响[D].牡丹江：牡丹江师范学院,2017.
[9]韩薇. α-突触核蛋白(SNCA)基因多态性与帕金森病易感性的meta分析[D].石家庄：河北医科大学,2015.
[10]PATIL K S,BASAK I,PAL R,et al. A proteomics approach to investigate miR1533p andmiR2055p targets in neuroblastoma cells[J]. PLoS ONE,2015,10(12):e0143969.
[11]DONG K,CHANG S,XIE Q,et al. Comparative transcriptomics of genetically divergent lines of chickens in response to Marek’s disease virus challenge at cytolytic phase[J]. PLoS ONE,2017,12(6):e0178923.
[12]GURUN A,KAMBLE N,KANFER B B,et al. Association of Marek’s disease induced immunosuppression with activation of a novel regulatory T cells in chickens [J]. PLoS Pathogens,2017,13(12):e1006745.
[13]PALMERO E I,CAMPOS M. Mechanisms and role of microRNA deregulation in cancer onset and progression[J]. Genetics and Molecular Biology,2011,34(3):363-370.
[14]YAO Y,ZHAO Y,SMITH L P,et al. Differential expression of microRNAs in Marek’s disease virustransformed Tlymphoma cell lines[J]. Journal of General Virology,2009,90(7): 1551-1559.
[15]TIAN F,LUO J,ZHANG H,et al. miRNA expression signatures induced by Marek’s disease virus infection in chickens[J]. Genomics,2012,99(3): 152-159.
[16]CORDES K R,SHEEHY N T,WHITE M P,et al. miR145 and miR143 regulate smooth muscle cell fate and plasticity[J]. Nature,2009,460:350-356.
[17]孙国荣. MDV分离毒株生物学特性及其在宿主染色体整合的研究[D].北京：中国农业科学院,2017.
[18]JUNN E,LEE K W,JEONG B S,et al. Repression of αsynuclein expression and toxicity by MicroRNA7[J]. Proceedings of the National Academy of Sciences of the United States of America,2009,106(31): 13052.

备注/Memo

备注/Memo:
收稿日期：2018-11-19 基金项目：国家自然科学基金项目（31500997） 作者简介：肖春婷（1992-）,女,贵州六盘水人,硕士研究生,研究方向为动物免疫遗传学。(E-mail)1063989121@qq.com 通讯作者：陈洪岩,(E-mail)sydw2014@163.com；廉传江,(E-mail)lcj121916@163.com
更新日期/Last Update: 2019-08-31