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家蚕中肠特异启动子BmAPN的克隆及活性分析
引用本文:陆改,程廷才,蒋亮,金盛凯,林平,胡翠美,夏庆友. 家蚕中肠特异启动子BmAPN的克隆及活性分析[J]. 中国农业科学, 2012, 45(20): 4279-4287. DOI: 10.3864/j.issn.0578-1752.2012.20.018
作者姓名:陆改  程廷才  蒋亮  金盛凯  林平  胡翠美  夏庆友
作者单位:1.西南大学蚕学与系统生物学研究所/家蚕基因组生物学国家重点实验室,重庆 400716
基金项目:国家“973”项目,国家“863”项目,国家自然科学基金
摘    要:【目的】克隆和鉴定1个家蚕中肠特异启动子,为研究家蚕的免疫应答和应用研究提供新的工具。【方法】从家蚕品种大造中提取基因组总DNA,用PCR方法克隆家蚕中肠特异表达基因BmAPN(GenBank登录号:BAA33715.1)的上游调控序列。利用此序列构建以EGFP为报告基因的转基因表达载体pBac[BmAPN-EGFP-SV40,3×P3-DsRed],通过显微注射获得转基因家蚕,在个体水平上检测启动子的活性。【结果】所克隆的BmAPN启动子长度为1 598 bp,其驱动的EGFP仅在转基因家蚕中肠特异表达,与内源基因BmAPN的表达特征一致。该启动子在家蚕幼虫时期活性相对较高,且在起蚕期的活性要明显高于眠蚕期,推测该启动子中的特异元件受激素调控。【结论】经克隆获得的BmAPN启动子为有活性的启动子,且为家蚕中肠特异启动子。

关 键 词:家蚕  中肠特异  启动子  转基因  
收稿时间:2012-02-13

Cloning and Activity Analysis of a Midgut-Specific Promoter BmAPN in Silkworm (Bombyx mori)
LU Gai , CHENG Ting-cai , JIANG Liang , JIN Sheng-kai , LIN Ping , HU Cui-mei , XIA Qing-you. Cloning and Activity Analysis of a Midgut-Specific Promoter BmAPN in Silkworm (Bombyx mori)[J]. Scientia Agricultura Sinica, 2012, 45(20): 4279-4287. DOI: 10.3864/j.issn.0578-1752.2012.20.018
Authors:LU Gai    CHENG Ting-cai    JIANG Liang    JIN Sheng-kai    LIN Ping    HU Cui-mei    XIA Qing-you
Affiliation:(Institute of Sericulture and Systems Biology,Southwest University/State Key Laboratory of Silkworm Genome Biology,Chongqing 400716)
Abstract:【Objective】The objective of this study is to provide a new tool for the immune-response and application research, and to clone and identify a midgut-specific promoter from Bombyx mori.【Method】 The 5′ upstream regulatory sequence of midgut-specific gene BmAPN (GenBank accession: BAA33715.1) was obtained by PCR from B. mori genomic DNA. A transgenic expression vector named pBac[BmAPN-EGFP-SV40, 3×P3-DsRed] was constructed, in which the reporter gene EGFP was driven by the BmAPN promoter. Furthermore, transgenic B. mori lines was obtained by microinjection and the activity of the promoter at the individual level were detected.【Result】The EGFP driven by a 1 598 bp BmAPN promoter was just specifically expressed in B. mori midgut. It was consistent with the expression pattern of endogenous gene BmAPN. The promoter activity was higher in B. mori larvae than other stages. And the activity in newly exuviated larvae was higher than that of molting larvae. It was speculated that specific components of the promoter might be regulated by hormone.【Conclusion】The BmAPN promoter is an active and midgut-specific promoter in B. mori.
Keywords:Bombyx mori  midgut-specific  promoter  transgene
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