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用于控制澳大利亚绵羊狂蝇 Lucilia cuprina 的遗传的转基因性别鉴定系统。

Transgenic sexing system for genetic control of the Australian sheep blow fly Lucilia cuprina.

机构信息

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

出版信息

Insect Biochem Mol Biol. 2014 Aug;51:80-8. doi: 10.1016/j.ibmb.2014.06.001. Epub 2014 Jun 11.

Abstract

The New World screwworm and the Australian sheep blowfly Lucilia cuprina are devastating pests of livestock. The larvae of these species feed on the tissue of the living animal and can cause death if untreated. The sterile insect technique or SIT was used to eradicate screwworm from North and Central America. This inspired efforts to develop strains containing complex chromosomal rearrangements for genetic control of L. cuprina in Australia. Although one field trial was promising, the approach was abandoned due to costs and difficulties in mass rearing the strain. As the efficiency of SIT can be significantly increased if only sterile males are released, we have developed transgenic strains of L. cuprina that carry a dominant tetracycline repressible female lethal genetic system. Lethality is due to overexpression of an auto-regulated tetracycline repressible transactivator (tTA) gene and occurs mostly at the pupal stage. Dominant female lethality was achieved by replacing the Drosophila hsp70 core promoter with a Lucilia hsp70 core promoter-5'UTR for tTA overexpression. The strains carry a dominant strongly expressed marker that will facilitate identification in the field. Interestingly, the sexes could be reliably sorted by fluorescence or color from the early first instar larval stage as females that overexpress tTA also overexpress the linked marker gene. Male-only strains of L. cuprina developed in this study could form the basis for a future genetic control program. Moreover, the system developed for L. cuprina should be readily transferrable to other major calliphorid livestock pests including the New and Old World screwworm.

摘要

新世界螺旋蝇和澳大利亚绵羊麻蝇 Lucilia cuprina 是对牲畜具有破坏性的害虫。这些物种的幼虫以活体动物的组织为食,如果不进行治疗,可能导致死亡。不育昆虫技术(SIT)曾被用于从北美和中美洲根除螺旋蝇。这激发了人们开发含有复杂染色体重排的品系的努力,以实现澳大利亚 L. cuprina 的遗传控制。尽管一次田间试验很有希望,但由于成本和大规模繁殖该品系的困难,该方法被放弃。由于如果只释放不育雄性,SIT 的效率可以显著提高,因此我们已经开发出携带显性四环素可抑制雌性致死遗传系统的转基因 L. cuprina 品系。致死性是由于自动调节的四环素可抑制转录激活剂(tTA)基因的过表达引起的,主要发生在蛹期。通过用 Lucilia hsp70 核心启动子-5'UTR 替换 Drosophila hsp70 核心启动子来实现 tTA 的过表达,从而实现显性雌性致死性。该品系携带一个显性强表达标记,这将有助于在现场识别。有趣的是,通过荧光或颜色可以从早期第一龄幼虫阶段可靠地对雌雄进行分类,因为过表达 tTA 的雌性也过表达连接的标记基因。本研究中开发的雄性-only L. cuprina 品系可以为未来的遗传控制计划奠定基础。此外,为 L. cuprina 开发的系统应该很容易转移到其他主要的 Calliphorid 牲畜害虫,包括新世界和旧世界螺旋蝇。

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