3×P3-EGFP marker facilitates screening for transgenic silkworm Bombyx mori L. from the embryonic stage onwards

Thomas, Jean-luc; Rocha, Martine Da; Besse, Arnaud; Mauchamp, Bernard; Chavancy, Gérard

doi:10.1016/s0965-1748(01)00150-3

Cited by 114 publications

(103 citation statements)

References 15 publications

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“…A transgenic technique was recently established in the silkworm, 7) and has been improved in marker gene 8) and expression 9) methods. The transgenic technique has been used in the functional analysis of genes 10,11) and as a tool in foreign gene production.…”

mentioning

confidence: 99%

A New Method for the Modification of Fibroin Heavy Chain Protein in the Transgenic Silkworm

Kojima¹,

Kuwana²,

Sezutsu³

et al. 2007

Bioscience, Biotechnology, and Biochemistry

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mentioning

confidence: 99%

A New Method for the Modification of Fibroin Heavy Chain Protein in the Transgenic Silkworm

Kojima¹,

Kuwana²,

Sezutsu³

et al. 2007

Bioscience, Biotechnology, and Biochemistry

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“…In the present study, we generated activator lines (GAL4 lines) in which GAL4 expression is under control of the B. mori cytoplasmic actin promoter (BmA3) (16) and effector lines (UAS lines) that carry the B. mori JHE gene (BmJHE) (11) linked to the UAS. The GAL4 and UAS lines also expressed the marker genes, DsRed2 (human codon optimized red fluorescent protein) and ECFP (enhanced cyan fluorescent protein) (18), under the control of the artificial eye-specific 3xP3 promoter, which allows for the identification of transgenic animals and further investigation from the embryonic stage (19,20). JHE was expressed at massively high levels in the progeny of the cross between the UAS and GAL4 lines.…”

mentioning

confidence: 99%

Precocious metamorphosis in transgenic silkworms overexpressing juvenile hormone esterase

Tan

Tanaka

Tamura

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

163

109

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Insect growth and development are intricately regulated by the titers of juvenile hormones (JHs) and ecdysteroids (and͞or their metabolites) in the insect hemolymph. Hydrolysis of the methyl ester of JH by a JH-specific esterase (JHE) is a key pathway for the degradation of JH. Here, we generate transgenic silkworm strains that overexpress JHE by using the binary GAL4͞UAS system. Overexpression of JHE from the embryonic stage resulted in larval-pupal metamorphosis after the third stadium, two stadia earlier than that observed in wild-type insects. This precocious metamorphosis suggests that JHs are not critical for normal development of embryo or larva before the second molt in Lepidoptera (moths and butterflies). Our transgenic approach allowed us to dissect the function of key physiological events that occur from embryogenesis. Until now, these types of studies were possible only in later larval stadia by using physical techniques such as allatectomy or the application of JH analogues. We believe that our system will allow further pioneering studies in insect physiology.Bombyx mori ͉ juvenile hormones ͉ transgenesis T wo major groups of nonpeptidic hormones, juvenile hormones (JHs) and ecdysteroids, play critical roles in the regulation of insect growth and development (1). The presence of JH during exposure to the ecdysteroids ensures a molt to a like stage, whereas the absence of JH during an ecdysteroid pulse allows metamorphosis. Concentrations of both hormones are regulated precisely by biosynthesis and degradation. Two definitive primary pathways for JH metabolism, hydrolysis of the methyl ester of JH by soluble esterases and hydration of the epoxide by microsomal epoxide hydrolases, have been described so far (2-5). In several insect orders, the key route of JH metabolism is by means of the hydrolysis of the methyl ester, and JH esterase (JHE) is considered the principal enzyme responsible for this function (2). Because of their importance in insect development, JHEs have been isolated and characterized from many insect orders such as Coleoptera (6), Diptera (7), and Lepidoptera (8-10), including the silkworm, Bombyx mori (11).As the primary degradative enzyme of JH, JHE has been used as an attractive tool for the study of JH action. Inhibition of JHE activity by a variety of 3-substituted 1,1,1-trifluoropropanone sulfides reduces the rate of JH degradation, thus delaying metamorphosis and resulting in giant larvae of Manduca sexta (12). By contrast, recombinant baculoviruses expressing JHE can reduce JH levels in Lepidoptera (13). However, difficulties in establishing stable and prompt in vivo expression of JHE in Lepidoptera have halted further studies.The mechanism of JH action in insects still remains a mystery despite a great deal of knowledge about its biological actions (5). Using Drosophila as a model for the study of JH action meets with difficulties because, in contrast to most insects, including those in Lepidoptera, Drosophila displays few morphogenetic actions that can be easily bioassayed ...

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“…Thus, under the A3 promoter, it is not possible to screen for transgenic individuals prior to G1 larvae. However, EGFP can be observed in the eggs of the G0 generation of Spodoptera litura, which can be used as screening vouchers [14].…”

Section: Comparison Of Transgenic Spodoptera Litura and Silkwormmentioning

confidence: 99%

Expression of pBac 3xP3 EGFP and pBac A3 EGFP Vector in Transgenic Spodoptera litura Fabricius

Gong

2018

TGE

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Abstract:In this study, the actin A3 promoter was inserted into piggyBac and constructed into pBacA3, which was inserted into piggyBac. The promoter was composed of the promoter and nerve-specific 3xP3 promoter, the enhanced green fluorescent protein (EGFP) gene and the SV40 polyadenylation recognition sequence. EGFP and pBac3xP3 EGFP transposon were injected into the early fertilized eggs of Spodoptera litura to detect whether they were expressed. The results showed that both vectors could be expressed in moth eggs, and positive individuals were obtained. The results showed that the expression rate of pBacA3 EGFP was higher than that of pBac3xP3 EGFP, and the proportion of EGFP in hatching larvae was higher than that in the latter. The results showed that pBacA3 EGFP was more suitable for Spodoptera litura Transposable carrier. In vitro transient expression of the transgenic vector of Spodoptera litura was not only the first step necessary to successfully carry out the transgenic gene of Spodoptera litura, but also itself can be applied to the study of gene function, which laid the foundation for the genome research of Spodoptera litura.

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3×P3-EGFP marker facilitates screening for transgenic silkworm Bombyx mori L. from the embryonic stage onwards

Cited by 114 publications

References 15 publications

A New Method for the Modification of Fibroin Heavy Chain Protein in the Transgenic Silkworm

A New Method for the Modification of Fibroin Heavy Chain Protein in the Transgenic Silkworm

Precocious metamorphosis in transgenic silkworms overexpressing juvenile hormone esterase

Expression of pBac 3xP3 EGFP and pBac A3 EGFP Vector in Transgenic Spodoptera litura Fabricius

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