Promoter and piggyBac activities within embryos of the potato tuber moth, Phthorimaea operculella, Zeller (Lepidoptera: Gelechiidae)

Mohammed, Ahmed; Coates, Craig J.

doi:10.1016/j.gene.2004.08.008

Cited by 16 publications

(20 citation statements)

References 31 publications

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“…First, based upon several successful excision assays that showed precise excision of piggyBac and repair of the TTAA target site in the donor plasmid, the piggyBac transposon is mobile in codling moth embryos. This assay has been a reliable predictor of transformation for Hobo, Hermes, and piggyBac transposons (Mohammed and Coates 2004;Raphael et al 2004;Lobo et al 1999;Michel et al 2001). Second, the evidential EGFP transient expression verifies that at least four promoters of EGFP work in the codling moth nuclear milieu: the Drosophila hsp70 promoter, the actin 5C promoter from D. melanogaster, the B. mori actin 3 promoter.…”

Section: Discussionmentioning

confidence: 78%

“…This is the sixth report of gene transfer in lepidopteran insects after the corn earworm, Helicoverpa zea (DeVault et al 1996); the pink bollworm, P. gossypiella (Peloquin et al 2000); the silkworm, B. mori (Tamura et al 2000); the potato tuber moth, Phthorimaea operculella (Mohammed and Coates 2004); and the butterfly Bicyclus anynana (Marcus et al 2004). This is the first report of a genetic transformation system for the codling moth.…”

Section: Discussionmentioning

confidence: 97%

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Genetic transformation of the codling moth, Cydia pomonella L., with piggyBac EGFP

et al. 2010

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Genetic transformation of the codling moth, Cydia pomonella, was accomplished through embryo microinjection with a plasmid-based piggyBac vector containing the enhanced green fluorescent protein (EGFP) gene. Sequencing of the flanking regions around the inserted construct resulted in identification of insect genomic sequences, not plasmid sequences, thus providing evidence that the piggyBac EGFP cassette had integrated into the codling moth genome. EGFP-positive moths were confirmed in the 28th and earlier generations post injection through PCR and Southern blot analyses, indicating heritability of the transgene.

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Section: Discussionmentioning

confidence: 78%

Section: Discussionmentioning

confidence: 97%

Genetic transformation of the codling moth, Cydia pomonella L., with piggyBac EGFP

et al. 2010

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“…Inclusion of a third plasmid expressing a transactivator is known to improve protein expression levels by enhancing promoter activity [30] or by increasing transpositional frequency of the element [29]. No significant variation in Gag expression was observed when the IE1 transactivator was included at transfection.…”

Section: Resultsmentioning

confidence: 99%

“…A transactivator plasmid (hr3IE1) to be included as a third plasmid in transfection, was supplied by K.J. Maragathavally [29,30]. Constructs harboring different regulatory elements were obtained from H. Zieler (see table 1) [31,32].…”

Section: Methodsmentioning

confidence: 99%

“…Two different helper plasmids, pCasper and pBSII-IE1, were tested for differences in transgene integration as assessed by subsequent Gag expression levels. The IE1 transactivator (hr3IE1) was included during transfection at a tenth of the total DNA amount to determine its potential to improve promoter activity or transposition frequency [29,30]. A comparison of transformability and transgenic Gag expression levels was conducted between three different insect cell lines, Sf21, Sf9 and High 5™.…”

Section: Methodsmentioning

confidence: 99%

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Use of the piggyBac transposon to create HIV-1 gag transgenic insect cell lines for continuous VLP production

et al. 2010

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BackgroundInsect baculovirus-produced Human immunodeficiency virus type 1 (HIV-1) Gag virus-like-particles (VLPs) stimulate good humoral and cell-mediated immune responses in animals and are thought to be suitable as a vaccine candidate. Drawbacks to this production system include contamination of VLP preparations with baculovirus and the necessity for routine maintenance of infectious baculovirus stock. We used piggyBac transposition as a novel method to create transgenic insect cell lines for continuous VLP production as an alternative to the baculovirus system.ResultsTransgenic cell lines maintained stable gag transgene integration and expression up to 100 cell passages, and although the level of VLPs produced was low compared to baculovirus-produced VLPs, they appeared similar in size and morphology to baculovirus-expressed VLPs. In a murine immunogenicity study, whereas baculovirus-produced VLPs elicited good CD4 immune responses in mice when used to boost a prime with a DNA vaccine, no boost response was elicited by transgenically produced VLPs.ConclusionTransgenic insect cells are stable and can produce HIV Pr55 Gag VLPs for over 100 passages: this novel result may simplify strategies aimed at making protein subunit vaccines for HIV. Immunogenicity of the Gag VLPs in mice was less than that of baculovirus-produced VLPs, which may be due to lack of baculovirus glycoprotein incorporation in the transgenic cell VLPs. Improved yield and immunogenicity of transgenic cell-produced VLPs may be achieved with the addition of further genetic elements into the piggyBac integron.

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Transposable Elements as Tools for Reshaping the Genome: It Is a Huge World After All!

Bire

Rouleux‐Bonnin

2012

Methods in Molecular Biology

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Promoter and piggyBac activities within embryos of the potato tuber moth, Phthorimaea operculella, Zeller (Lepidoptera: Gelechiidae)

Cited by 16 publications

References 31 publications

Genetic transformation of the codling moth, Cydia pomonella L., with piggyBac EGFP

Genetic transformation of the codling moth, Cydia pomonella L., with piggyBac EGFP

Use of the piggyBac transposon to create HIV-1 gag transgenic insect cell lines for continuous VLP production

Transposable Elements as Tools for Reshaping the Genome: It Is a Huge World After All!

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