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当前位置: 首页 > 文章 > Bmo-miR-2788对家蚕丝胶蛋白基因Ser-1的表达调控（英文） 蚕业科学 2019 (6) 811-820
Position: Home > Articles > Regulation on Expression of Bombyx mori Sericin-1 Gene by BmomiR-2788 Science of Sericulture 2019 (6) 811-820

Bmo-miR-2788对家蚕丝胶蛋白基因Ser-1的表达调控（英文）

作  者：
Rehana Kandhro;陈艳花;钱平;朱娟;沈兴家
单  位：
中国农业科学院蚕业研究所农业农村部蚕桑遗传改良重点实验室;江苏科技大学生物技术学院江苏省蚕桑生物学与生物技术重点实验室
关键词：
miRNA;BmN细胞;丝胶蛋白基因;BmSer-1;Bmo-miR-2788;基因调控
摘  要：
通过生物信息学分析发现,家蚕丝胶蛋白基因BmSer-1的3′非翻译区(3′-UTR)存在微小RNA Bmo-miR-2788的结合位点。半定量PCR结果显示,Bmo-miR-2788在家蚕5龄第4天幼虫的8个受试组织中均有表达,但在中部丝腺中表达水平最高;而BmSer-1仅在中部丝腺中被检测到。分别构建BmSer-1 3′-UTR和Bmo-miR-2788的表达载体pGL3.0(A3-luc-BmSer-1-3′-UTR-SV40)和pcDNA3.0(ie1-egfp-pri-Bmo-miR-2788-SV40),共转染BmN细胞,结果表明Bmo-miR-2788上调BmSer-1表达(P<0.01);当将上述载体与合成的Bmo-miR-2788抑制剂共转染BmN细胞时,BmSer-1表达显著下调(P<0.05)。为验证Bmo-miR-2788在蚕体内对BmSer-1表达的调控作用,将Bmo-miR-2788表达质粒与其阴性对照(pcDNA3.0)分别注射到家蚕5龄第3天幼虫体内,qRT-PCR分析结果显示,与阴性对照相比,注射Bmo-miR-2788后36 h中部丝腺中的BmSer-1表达显著上调(P<0.01)。上述结果表明Bmo-miR-2788正向调控BmSer-1基因的表达。
译  名：
Regulation on Expression of Bombyx mori Sericin-1 Gene by BmomiR-2788
作  者：
Rehana Kandhro;Chen Yanhua;Qian Ping;Zhu Juan;Shen Xingjia;Jiangsu Key Laboratory of Sericutural Biology and Biotechnology,College of Biotechnology,Jiangsu University of Science and Technology;Key Laboratory of Silkworm and Mulberry Genetic Improvement,Ministry of Agriculture and Rural Affairs,Sericultural Research Institute,Chinese Academy of Agricultural Sciences;
关键词：
miRNA;;BmN cell;;Sericin gene;;BmSer-1;;Bmo-miR-2788;;Gene regulation
摘  要：
Integrated bioinformatics analysis revealed that there was a binding site for miRNA Bmo-miR-2788 in the 3′-untranslated region(3′-UTR)of sericin gene BmSer-1. Semi-quantitative polymerase chain reaction(PCR)results showed that Bmo-miR-2788 was expressed in 8 tissues of Bombyx mori larvae of 4 th day 5 th instar,and the highest expression was detected in the middle silk gland(MSG). BmSer-1 expression was detected only in the MSG. Then,expression vectors of BmSer-1 3′-UTR and Bmo-miR-2788 were constructed. After pGL3. 0(A3-luc-BmSer-1-3′-UTR-SV40)and pcDNA3. 0(ie1-egfp-pri-Bmo-miR-2788-SV40)were co-transfected in BmN cells,BmSer-1 expression was up-regulated by Bmo-miR-2788(P<0. 01). When the above vectors and synthetic Bmo-miR-2788 inhibitor were co-transfected in BmN cells,BmSer-1 expression was significantly down-regulated(P<0. 05). Additionally,Bmo-miR-2788 expressed plasmid and its negative control(pcDNA3. 0)were injected into 3 rd day 5 th instar silkworm larvae to validate the function of BmomiR-2788 on BmSer-1 expression in vivo. Quantitative real-time PCR(qRT-PCR)results showed that Bmo-miR-2788 significantly up-regulated BmSer-1 expression in MSG 36 h post injection(P<0. 01)as compared with the negative control.All above results demonstrate that Bmo-miR-2788 can positively regulate expression of BmSer-1 gene.

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