Lobo N, Li X, Hua-Van A, Fraser M J
Department of Biology, University of Notre Dame, IN 46556, USA.
Mol Genet Genomics. 2001 Mar;265(1):66-71. doi: 10.1007/s004380000388.
The re-emergence of arboviral diseases such as Dengue Fever and La Crosse encephalitis is primarily due to the failure of insect vector control strategies. The development of a procedure capable of producing stable germ-line transformants in the insect vectors of these diseases would bridge the gap between gene expression systems being developed to curb vector transmission and the identification of important genes and regulatory sequences and their reintroduction back into the insect genome in the form of vector control strategies. The transposable element piggyBac is capable of transposition in a variety of insect species, and could serve as a versatile insect transformation vector. Using plasmid-based excision and transposition assays, we report that this short-ITR transposon undergoes precise, transposase-dependent excision and transposition in embryos of Aedes albopictus and Aedes triseriatus, the vectors of Dengue fever and LaCrosse encephalitis, respectively. These assays allow us easily and rapidly to confirm and assess the potential utility of piggyBac as a gene transfer tool in a given species. piggyBac is an exceptionally mobile and versatile genetic transformation vector, comparable to other transposons currently in use for the transformation of insects. The mobility of the piggyBac element seen in both Ae. albopictus and Ae. triseriatus is further evidence that it can be employed as a germ-line vector in important insect disease vectors.
登革热和拉克罗斯脑炎等虫媒病毒疾病的再度出现主要是由于昆虫媒介控制策略的失败。开发一种能够在这些疾病的昆虫媒介中产生稳定种系转化体的方法，将弥合为遏制媒介传播而开发的基因表达系统与重要基因和调控序列的鉴定之间的差距，并将它们以媒介控制策略的形式重新引入昆虫基因组。转座元件piggyBac能够在多种昆虫物种中进行转座，可作为一种通用的昆虫转化载体。通过基于质粒的切除和转座分析，我们报告这种短末端反向重复（short-ITR）转座子在白纹伊蚊和三带喙库蚊（分别是登革热和拉克罗斯脑炎的媒介）的胚胎中进行精确的、转座酶依赖性的切除和转座。这些分析使我们能够轻松快速地确认和评估piggyBac作为给定物种中基因转移工具的潜在效用。piggyBac是一种异常活跃且通用的遗传转化载体，可与目前用于昆虫转化的其他转座子相媲美。在白纹伊蚊和三带喙库蚊中观察到的piggyBac元件的移动性进一步证明它可作为重要昆虫疾病媒介中的种系载体。
