Gga-miR-7b和靶基因SNCA在两品系鸡脾脏中的表达分析  

Expression analysis of Gga-miR-7b and target gene SNCA in spleen of two strains of chicken

作　　者：肖春婷 廉传江[2] 易诚 陈洪岩[1,2] XIAO Chun-ting;LIAN Chuan-jiang;YI Cheng;CHEN Hong-yan(College of Life Science, Northeast Agricultural University, Harbin 150030, China;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;College of Life Science, Northeast Forestry University, Harbin 150040, China)

机构地区：[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

出　　处：《江苏农业学报》2019年第4期874-879,共6页Jiangsu Journal of Agricultural Sciences

基　　金：国家自然科学基金项目(31500997)

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。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.

关 键 词：gga-miR-7b SNCA 靶向关系 表达分析 

分 类 号：Q786[生物学—分子生物学]