Efficient germ-line transformation of the economically important pest species Lucilia cuprina and Lucilia sericata (Diptera, Calliphoridae)

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.

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► A strong marker gene developed using the constitutive Lucilia cuprina hsp83 (Lchsp83) gene promoter to drive expression of the ZsGreen fluorescent protein gene. Germ-line transformation efficiency of L. cuprina significantly improved by using a piggyBac vector containing the Lchsp83-ZsGreen marker and a piggyBac helper that used the Lchsp83 promoter to control piggyBac transposase gene expression. Most transgenic lines contain a single copy of the piggyBac transgene. First report of transformation of L. sericata using the efficient piggyBac vector/helper combination.