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.
Concha
Carolina
C
Institute of Molecular BioSciences, Massey University, Private Bag 11222, Palmerston North, New Zealand. concha@cict.fr
Belikoff
Esther J
EJ
Carey
Brandi-lee
BL
Li
Fang
F
Schiemann
Anja H
AH
Scott
Maxwell J
MJ
D000818:Animals; D030801:Animals, Genetically Modified / Q000378:metabolism*; D001315:Australia; D004175:Diptera / Q000235:genetics*; D005260:Female; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D017344:Genes, Insect; D005854:Germ Cells / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000032:analysis; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007814:Larva / Q000235:genetics / Q000378:metabolism; D009520:New Zealand; D010572:Pest Control, Biological / Q000379:methods; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D012756:Sheep; D014170:Transformation, Genetic; D019895:Transposases / Q000235:genetics / Q000378:metabolism; D014947:Wounds and Injuries / Q000628:therapy
D006360:Heat-Shock Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D019895:Transposases