Institute of Molecular BioSciences, Massey University, Private Bag 11222, Palmerston North, New Zealand.
Insect Biochem Mol Biol. 2011 Jan;41(1):70-5. doi: 10.1016/j.ibmb.2010.09.006. Epub 2010 Sep 29.
The green blowfly species Lucilia cuprina and Lucilia sericata are economically important pests for the sheep industries of Australia and New Zealand. L. cuprina has long been considered a good target for a genetic pest management program. In addition, L. sericata maggots are used in the cleaning of wounds and necrotic tissue of patients suffering from ulcers that are difficult to treat by other methods. Development of efficient transgenesis methods would greatly facilitate the development of strains ideal for genetic control programs or could potentially improve "maggot therapy". We have previously reported the germ-line transformation of L. cuprina and the design of a "female killing system" that could potentially be applied to this species. However, the efficiency of transformation obtained was low and transformed lines were difficult to detect due to the low expression of the EGFP marker used. Here we describe an efficient and reliable method for germ-line transformation of L. cuprina using new piggyBac vector and helper plasmids containing the strong promoter from the L. cuprina hsp83 gene to drive expression of the transposase and fluorescent protein marker gene. We also report, for the first time, the germ-line transformation of L. sericata using the new piggyBac vector/helper combination.
绿头苍蝇属的物种 Lucilia cuprina 和 Lucilia sericata 是澳大利亚和新西兰绵羊产业中具有重要经济意义的害虫。L. cuprina 长期以来一直被认为是遗传害虫管理计划的一个很好的目标。此外，L. sericata 蛆虫被用于清洁溃疡患者的伤口和坏死组织，这些溃疡用其他方法难以治疗。开发高效的转基因方法将极大地促进适合遗传控制计划的菌株的发展，或者有可能改善“蛆虫疗法”。我们之前已经报道了 L. cuprina 的种系转化，以及一种“雌性致死系统”的设计，该系统可能适用于该物种。然而，由于使用的 EGFP 标记表达水平低，转化效率低，转化系难以检测。在这里，我们描述了一种使用新的 piggyBac 载体和含有 L. cuprina hsp83 基因强启动子的辅助质粒的高效可靠的 L. cuprina 种系转化方法，以驱动转座酶和荧光蛋白标记基因的表达。我们还首次报道了使用新的 piggyBac 载体/辅助组合对 L. sericata 的种系转化。
