Hermes‐mediated germ‐line transformation of the Mediterranean fruit fly Ceratitis capitata

Abstract

We report the use of the Hermes transposable element for germ-line transformation of the Mediterranean fruit fly, Ceratitis capitata. Hermes was able to genetically transform this insect at an estimated frequency between 0.6 and 1.1%, which is comparable to the transformation frequencies obtained for this species when using other transposable elements. Hermes integrates into the medfly genome by a cut-and-paste mechanism and the sequences integrated into the genome are delimited by the terminal nucleotides of the Hermes inverted terminal repeats. Integration resulted in the generation of 8 bp target site duplications, the sequences of which conformed to the target site duplications generated by hAT element transposition in insects. The Hermes element is one additional genetic tool that can be deployed in manipulating and characterizing the medfly genome.

Current Trends in Quarantine Entomology

“…The term, originally coined by Fryxell & Miller (35), was based on an idea from Knipling's group (77), in which they proposed to develop a nondiapausing strain of Diabrotica and perform overflooding releases in a type of self-killing (autocidal) pest control program. Genetically engineering insects using transposable elements such as Hobo, piggyBac (17,43,44,56), Hermes (89,104), and minos (83) facilitates strain development. With these high-efficiency transposable elements available for insect transformation, it has been proposed that pest insects can be transformed with conditionally lethal genes that would be controlled under laboratory conditions to allow for mass rearing and become lethal under normal environmental conditions (35).…”

Gene vector and transposable element behavior in mosquitoes

“…These alternate forms of the right ITR, referred to as B5 and B6, involve a G-to-C transversion in the terminal nucleotide and originated during the isolation of two independent genomic DNA fragments from Musca domestica containing a Hermes element (Warren et al, 1994;Fig.·1). Elements containing the B5 end (with a terminal G) have been used in most of the studies of Hermes in insects (Allen et al, 2001;Michel et al, 2001;Pinkerton et al, 1996Pinkerton et al, , 2000Sarkar et al, 1997a,b), while the element containing the B6 ITR (with a terminal C) has been used in A. aegypti transformation experiments reported by others Kokoza et al, 2000;Moreira et al, 2000).…”

DNA sequence requirements for hobo transposable element transposition in Drosophila melanogaster

“…The truncated ORF1 was removed from pBShoboATG3 as a 1.8 kb Ecl136 II-Hpa I fragment, which was then inserted into the Hinc II site of pKhsp82 yielding pKhsp82hoboATG3. The truncated ORF1 was placed downstream of the hsp70 promoter in pKhsp70 (Michel et al 2001). Plasmid pBShoboATG3 was cleaved with Ecl136 II and Kpn I, which was blunt-ended by T4 DNA polymerase and cloned into the Sma I site of pKhsp70 to generate pKhsp70hoboATG3.…”

“…Insect hAT elements, such as hobo, the Hermes element of Musca domestica, the Herves element of Anopheles gambiae and the recently discovered active Buster elements from Aedes aegypti and Tribolium castaneum appear to be widely distributed amongst insects and, in the case of Hermes, have proven to be useful transformation vectors of several non-drosophilid insect species Arensburger et al 2005Arensburger et al , 2011O'Brochta et al 1996O'Brochta et al , 2000Jasinskiene et al 1998;Michel et al 2001;Allen et al 2001;Marcus et al 2004). The insect hAT elements are unique in that, unlike the P, mariner, Minos, and piggyBac systems, they consist of five characterized elements that are active in host and non-host species (O'Brochta et al , 1996Arensburger et al 2005Arensburger et al , 2011.…”