Transgenic characterization of two testis‐specific promoters in the silkworm, Bombyx mori
Transgenic characterization of two testis‐specific promoters in the silkworm, Bombyx mori
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DOI：
10.1111/imb.12144
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发表时间：
2015-04
影响因子：
2.6
通讯作者：
Jun Xu;H. Bi;R. Chen;A. F. Aslam;Zhi-Qian Li;L. Ling;B. Zeng;Yongping Huang;A. Tan
中科院分区：
文献类型：
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作者：
Jun Xu;H. Bi;R. Chen;A. F. Aslam;Zhi-Qian Li;L. Ling;B. Zeng;Yongping Huang;A. Tan
性别特异性调控元件是开发昆虫遗传性别系统的关键组成部分。目前的昆虫遗传性别鉴定系统主要使用雌性特异性遗传修饰系统，而雄性特异性遗传修饰几乎没有成功的报道。在具有经济重要性的鳞翅目模式昆虫蚕 Bombyx mori 中，基于性别特异性选择性剪接因子建立了基于转基因的雌性特异性致死系统，并鉴定了雌性特异性启动子 BmVgp（卵黄蛋白原启动子）。然而，尚未确定男性特异性的调控元件。在这里，我们报告了两个启动子的转基因鉴定，这两个启动子在家蚕中以睾丸特异性方式驱动报告基因表达。使用基于 Piggybac 的种系转化引入来自家蚕径向辐条头 1 基因 (BmR1) 和 β-微管蛋白 4 基因 (Bmβ4) 的推定启动子序列。在转基因蚕中，由 BmR1 启动子 (BmR1p) 或 Bmβ4p 指导的报告基因增强型绿色荧光蛋白 (EGFP) 的表达显示出从幼虫到成虫阶段精确的睾丸特异性方式。此外，这两个转基因系的 EGFP 表达在睾丸中显示出不同的定位，表明 BmR1p 或 Bmβ4p 可能用作指导睾丸特异性基因表达的不同调控元件。这些睾丸特异性启动子的鉴定不仅有助于更好地了解昆虫睾丸特异性基因的功能，而且在害虫管理的昆虫不育技术中具有潜在的应用。
Sex‐specific regulatory elements are key components for developing insect genetic sexing systems. The current insect genetic sexing system mainly uses a female‐specific modification system whereas little success was reported on male‐specific genetic modification. In the silkworm Bombyx mori, a lepidopteran model insect with economic importance, a transgene‐based, female‐specific lethality system has been established based on sex‐specific alternative splicing factors and a female‐specific promoter BmVgp (vitellogenin promoter) has been identified. However, no male‐specific regulatory elements have yet been identified. Here we report the transgenic identification of two promoters that drive reporter gene expression in a testis‐specific manner in B. mori. Putative promoter sequences from the B. mori Radial spoke head 1 gene (BmR1) and beta‐tubulin 4 gene (Bmβ4) were introduced using piggybac‐based germline transformation. In transgenic silkworms, expression of the reporter gene enhanced green fluorescent protein (EGFP) directed by either BmR1 promoter (BmR1p) or Bmβ4p showed precisely testis‐specific manners from the larval to adult stage. Furthermore, EGFP expression of these two transgenic lines showed different localization in the testis, indicating that BmR1p or Bmβ4p might be used as distinct regulatory elements in directing testis‐specific gene expression. Identification of these testis‐specific promoters not only contributes to a better understanding of testis‐specific gene function in insects, but also has potential applications in sterile insect techniques for pest management.