Handler Alfred M
Center for Medical, Agricultural, and Veterinary Entomology, Agricultural Research Service, US Department of Agriculture, Gainesville, FL 32608, USA.
Genetica. 2002 Sep;116(1):137-49. doi: 10.1023/a:1020924028450.
The genetic manipulation of non-drosophilid insect species is possible by the creation of recombinant DNA constructs that can be integrated into host genomes by several transposon-based vector systems. This technology will allow the development and testing of a variety of systems that can improve existing biological control methods, and the development of new highly efficient methods. For programs such as sterile insect technique (SIT), transgenic strains may include fluorescent protein marker genes for detection of released insects, and conditional gene expression systems that will result in male sterility and female lethality for genetic sexing. Conditional expression systems include the yeast GAL4 system and the bacterial Tet-off and Tet-on systems that can, respectively, negatively or positively regulate expression of genes for lethality or sterility depending on a dietary source of tetracycline. Importantly, strains for male sterility must also incorporate an effective system for genetic sexing, since typically, surviving females would remain fertile. Models for the use of these expression systems and associated genetic material come from studies in Drosophila and, while many of these systems should be transferable to other insects, continued research will be necessary in insects of interest to clone genes, optimize germ-line transformation, and perform vector stability studies and risk assessment for their release as transgenic strains.
通过构建重组DNA构建体，利用几种基于转座子的载体系统将其整合到宿主基因组中，从而实现对非果蝇昆虫物种的基因操作。这项技术将有助于开发和测试各种系统，这些系统可以改进现有的生物防治方法，并开发新的高效方法。对于诸如昆虫不育技术（SIT）等项目，转基因品系可能包括用于检测释放昆虫的荧光蛋白标记基因，以及能导致雄性不育和雌性致死以进行遗传性别鉴定的条件性基因表达系统。条件性表达系统包括酵母GAL4系统以及细菌的Tet-off和Tet-on系统，它们可以分别根据四环素的饮食来源对致死或不育基因的表达进行负调控或正调控。重要的是，雄性不育品系还必须包含一个有效的遗传性别鉴定系统，因为通常情况下，存活的雌性仍会保持生育能力。使用这些表达系统及相关遗传物质的模型来自于果蝇研究，虽然其中许多系统应该可以转移到其他昆虫上，但对于感兴趣的昆虫，仍需要继续开展研究以克隆基因、优化种系转化，并对其作为转基因品系释放进行载体稳定性研究和风险评估。
