| 家蚕滞育激素受体体外上调下游海藻糖酶基因的表达 |
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| 引用本文： | 沈兴家,;王力刚,;韦博尤,;朱娟,;唐顺明,;赵巧玲.家蚕滞育激素受体体外上调下游海藻糖酶基因的表达[J].江苏科技大学学报(社会科学版),2014(3):298-302. |
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| 作者姓名： | 沈兴家 ;王力刚 ;韦博尤 ;朱娟 ;唐顺明 ;赵巧玲 |
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| 作者单位： | [1]江苏科技大学蚕业研究所,江苏镇江212018; [2]中国农业科学院蚕业研究所,江苏镇江212018; [3]广西蚕业科学研究院,广西南宁530007 |
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| 基金项目： | 国家自然科学基金资助项目（31172266） |
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| 摘 要： | 家蚕滞育激素（Bombyx mori diapause hormone，BmDH）与滞育激素受体（BmDH receptor，BmDHR）结合，调控其信号通路中的下游基因的表达.在二化性家蚕中，受滞育信号的影响，蛹体卵巢膜上海藻糖酶的活性增强.为了证实 BmDHR对家蚕海藻糖酶基因（Bmtre）表达的调控作用，以 pcDNA3.1为载体，构建了4种由 ie -1启动子驱动的不同 Bmdhr 的表达质粒 pcDNA - ie1- Bmdhr1、pcDNA - ie1- Bmdhr3、pcDNA - ie1- Bmdhr4、pcDNA - ie1- Bmdhr5和（Bmtre 启动子驱动的萤火虫荧光素酶基因（luciferase，luc）报告质粒 p3Z - Bmtre - luc；用脂质体（Lipofectin）包埋1种 Bmdhr 表达质粒或不同摩尔比的 pcDNA - ie1- Bmdhr1/ pcDNA - ie1- Bmdhr5以及报告质粒后，分别共转染源于卵巢的家蚕细胞 BmN，在 DH 作用下体外检测 BmDHRs 对海藻糖酶基因启动子活性的影响.结果表明，4种滞育激素受体 BmDHR -1、BmDHR -3、BmDHR -4和BmDHR -5分别表达时，均能上调海藻糖酶基因启动子的活性；不同摩尔比（1：3，1：1，3：1）的 BmDHR -1/ BmDHR -5也均能上调海藻糖酶基因启动子的活性，由此证明 BmDHR 能上调其信号通路中下游基因 Bmtre 的表达.
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| 关 键 词： | 家蚕 滞育激素受体 海藻糖酶 基因表达调控 |
Diapause hormone receptors of Bombyx mori up-regulate expression of its downstream gene trehalase in vitro |
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| Institution: | Shen Xingjia, Wang Ligang, Wei Boyou, Zhu Juan, Tang Sunming, Zhao Qiaoling (1. Sericultural Research Institute, Jiangsu University of Science and Techonology, Zhenjiang Jiangsu 212018, China) ( 2. Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang Jiangsu 212018, China) ( 3. Guangxi Academy of Sericultural Sciences, Nanning Guangxi 530007, China) |
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| Abstract: | Bombyx mori diapause hormone（DH）bonds with diapause hormone receptors（BmDHR）to regulate downstream genes at its signal passway. In the pupal stage of bivoltine silkworm B. mori,the enzyme activity of trehalase is enhanced by diapause signal on the membrane of ovaries. To validate the regulation function of Bm-DHR on Bombyx mori trehalase gene（Bmtre）,we constructed 4 different BmDHR expression plasmids driven by ie - 1 promoter by using pcDNA3. 1 vector,pcDNA - ie1 - Bmdhr1,pcDNA - ie1 - Bmdhr3,pcDNA - ie1 -Bmdhr4 and pcDNA - ie1 - Bmdhr5,and a report plasmid p3Z - Bmtre - luc with a luciferase gene driven by Bmtre promoter. Then BmN cells delivered from the ovary of B. mori were co - transfected respectively with above one BmDHR expression plasmid or different Mole ratio of BmDHR - 1 / BmDHR - 5,and reported plasmid medi-ated by lipofectin. The effects of BmDHR on the Bmtre promoter activity were detected respectively. The results showed that BmDHR - 1,BmDHR - 3,BmDHR - 4 and BmDHR - 5 increased the activity of Bmtre promoter independently. Different ratios of BmDHR - 1 / BmDHR - 5（1： 3,1： 1 and 3： 1）also increased Bmtre promoter activity. These results indicate that BmDHRs can up-regulate the expression of downstream gene Bmtre at its sig-nal passway. |
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| Keywords: | Bombyx mori diapause hormone receptor trehalase regulation of gene expression |
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