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一种用于澳大利亚羊蝇 Lucilia cuprina 的转基因胚胎性别鉴定系统。

A transgenic embryonic sexing system for the Australian sheep blow fly Lucilia cuprina.

作者信息

Yan Ying, Scott Maxwell J

机构信息

Department of Entomology, North Carolina State University, Campus Box 7613, Raleigh, NC, 27695-7613.

出版信息

Sci Rep. 2015 Nov 5;5:16090. doi: 10.1038/srep16090.

Abstract

Genetic approaches, including the sterile insect technique (SIT), have previously been considered for control of the Australian sheep blow fly Lucilia cuprina, a major pest of sheep. In an SIT program, females consume 50% of the diet but are ineffective as control agents and compete with females in the field for mating with sterile males, thereby decreasing the efficiency of the program. Consequently, transgenic sexing strains of L. cuprina were developed that produce 100% males when raised on diet that lacks tetracycline. However, as females die mostly at the pupal stage, rearing costs would not be significantly reduced. Here we report the development of transgenic embryonic sexing strains of L. cuprina. In these strains, the Lsbnk cellularization gene promoter drives high levels of expression of the tetracycline transactivator (tTA) in the early embryo. In the absence of tetracycline, tTA activates expression of the Lshid proapoptotic gene, leading to death of the embryo. Sex-specific RNA splicing of Lshid transcripts ensures that only female embryos die. Embryonic sexing strains were also made by combining the Lsbnk-tTA and tetO-Lshid components into a single gene construct, which will facilitate transfer of the technology to other major calliphorid livestock pests.

摘要

包括昆虫不育技术(SIT)在内的遗传方法,此前已被考虑用于控制澳大利亚羊绿蝇Lucilia cuprina,这是绵羊的一种主要害虫。在昆虫不育技术项目中,雌性消耗50%的食物,但作为防治媒介无效,且在野外与雌性竞争与不育雄性交配,从而降低了该项目的效率。因此,开发了转基因性别品系的绿蝇,当在缺乏四环素的食物上饲养时,该品系产生100%的雄性。然而,由于雌性大多在蛹期死亡,饲养成本不会显著降低。在此,我们报告了转基因胚胎性别品系绿蝇的开发。在这些品系中,Lsbnk细胞化基因启动子在早期胚胎中驱动四环素反式激活因子(tTA)的高水平表达。在没有四环素的情况下,tTA激活Lshid促凋亡基因的表达,导致胚胎死亡。Lshid转录本的性别特异性RNA剪接确保只有雌性胚胎死亡。通过将Lsbnk-tTA和tetO-Lshid组件组合成一个单一基因构建体,也制备了胚胎性别品系,这将便于将该技术转移到其他主要的丽蝇科家畜害虫。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/19d6/4633611/1d007d519da2/srep16090-f1.jpg

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