Scott Maxwell J, Atapattu Asela, Schiemann Anja H, Concha Carolina, Henry Rebecca, Carey Brandi-lee, Belikoff Esther J, Heinrich Jörg C, Sarkar Abhimanyu
Department of Genetics, North Carolina State University, Campus Box 7614, North Carolina, USA.
Genetica. 2011 Jan;139(1):63-70. doi: 10.1007/s10709-010-9492-6. Epub 2010 Sep 16.
The Australian sheep blowfly Lucilia cuprina is a major pest for the Australian and New Zealand sheep industries. With the long-term aim of making a strain of L. cuprina suitable for a genetic control program, we previously developed a tetracycline-repressible female lethal genetic system in Drosophila. A key part of this system is a female-specific promoter from a yolk protein (yp) gene controlling expression of the tetracycline-dependent transactivator (tTA). Here we report the sequence of a 14.2 kb genomic clone from L. cuprina that contains a cluster of three complete yp genes and one partial yp gene. The Lcyp genes are specifically expressed in females that have received a protein meal. A bioinformatic analysis of the promoter of one of the yp genes (LcypA) identified several putative binding sites for DSX, a known regulator of yp gene expression in other Diptera. A transgenic strain of L. cuprina was made that contained the LcypA promoter driving the expression of the Escherichia coli lacZ reporter gene. Transgenic females express high levels of β-galactosidase after a protein meal. Thus the LcypA promoter could be used to obtain female-specific expression of tTA in transgenic L. cuprina.
澳大利亚羊蝇 Lucilia cuprina 是澳大利亚和新西兰养羊业的主要害虫。为了培育出适合遗传控制计划的 Lucilia cuprina 品系，我们此前在果蝇中开发了一种四环素可抑制的雌性致死遗传系统。该系统的一个关键部分是来自卵黄蛋白（yp）基因的雌性特异性启动子，它控制四环素依赖性反式激活因子（tTA）的表达。在此，我们报告了来自 Lucilia cuprina 的一个 14.2 kb 基因组克隆的序列，该克隆包含一组三个完整的 yp 基因和一个部分 yp 基因。Lcyp 基因在摄取了蛋白质食物的雌性中特异性表达。对其中一个 yp 基因（LcypA）的启动子进行生物信息学分析，确定了几个可能的 DSX 结合位点，DSX 是其他双翅目昆虫中已知的 yp 基因表达调节因子。构建了一个含有驱动大肠杆菌 lacZ 报告基因表达的 LcypA 启动子的 Lucilia cuprina 转基因品系。转基因雌性在摄取蛋白质食物后表达高水平的β-半乳糖苷酶。因此，LcypA 启动子可用于在转基因 Lucilia cuprina 中实现 tTA 的雌性特异性表达。
