Transgene expression from the <i>Tribolium castaneum Polyubiquitin</i> promoter

Lorenzen, Marcé D.; Brown, Susan J.; Denell, R. E.; Beeman, Richard W.

doi:10.1046/j.1365-2583.2002.00349.x

Cited by 40 publications

(44 citation statements)

References 30 publications

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“…17 using the primers listed in Table 1, which is published as supporting information on the PNAS web site. The following primers designed for the Tribolium polyubiquitin gene (18) were used as an internal control for normalization of equal sample loading: 5Ј-GACCGGCAAGACCATCACT-3Ј and 5Ј-CGCAGACGCAAAACTGAAGG-3Ј. For TcLac2 RNAi, dsLac2 was injected into prepupae, and total RNA was isolated 5-6 d after pupation (6-7 d after injection).…”

Section: Analysis Of Expression By Rt-pcrmentioning

confidence: 99%

Laccase 2 is the phenoloxidase gene required for beetle cuticle tanning

Arakane

Muthukrishnan

Beeman

et al. 2005

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

339

343

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Cuticle tanning (or sclerotization and pigmentation) in invertebrates involves the oxidative conjugation of proteins, which renders them insoluble and hardens and darkens the color of the exoskeleton. Two kinds of phenoloxidases, laccase and tyrosinase, have been proposed to participate in tanning, but proof of the true identity of the enzyme(s) responsible for this process has been elusive. We report the cloning of cDNAs for laccases and tyrosinases from the red flour beetle, Tribolium castaneum, as well as their developmental patterns of expression. To test for the involvement of these types of enzymes in cuticle tanning, we performed RNA interference experiments to decrease the levels of individual phenoloxidases. Normal phenotypes were obtained after dsRNA-mediated transcript depletion for all phenoloxidases tested, with the exception of laccase 2. Insects injected with dsRNA for the laccase 2 gene failed to tan, were soft-bodied and deformed, and subsequently died in a dsRNA dose-dependent fashion. The results presented here support the hypothesis that two isoforms of laccase 2 generated by alternative splicing catalyze larval, pupal, and adult cuticle tanning in Tribolium.exoskeleton ͉ sclerotization ͉ pigmentation ͉ Tribolium castaneum ͉ RNA interference

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Section: Analysis Of Expression By Rt-pcrmentioning

confidence: 99%

Laccase 2 is the phenoloxidase gene required for beetle cuticle tanning

Arakane

Muthukrishnan

Beeman

et al. 2005

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

339

343

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“…Like Drosophila, Tribolium is amenable to genetic manipulation through germ line transformation, transgene expression vectors and RNAi; the genome has also been sequenced and assembled into linkage maps that are available online via Beetlebase (Browne and Scholtz, 1999;Lorenzen et al, 2002Lorenzen et al, , 2003Lorenzen et al, , 2005Wang et al, 2007). Unlike the fruit fly and similar to the cricket, the mushroom body neuroblasts of beetles persist into adulthood and undergo active neurogenesis as indicated by BrdU incorporation (Cayre et al, 1996).…”

Section: Introductionmentioning

confidence: 99%

Metamorphosis and adult development of the mushroom bodies of the red flour beetle, Tribolium castaneum

Zhao

Coptis

Farris

2008

Developmental Neurobiology

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The insect mushroom bodies play important roles in a number of higher processing functions such as sensory integration, higher level olfactory processing, and spatial and associative learning and memory. These functions have been established through studies in a handful of tractable model systems, of which only the fruit fly Drosophila melanogaster has been readily amenable to genetic manipulations. The red flour beetle Tribolium castaneum has a sequenced genome and has been subject to the development of molecular tools for the ready manipulation of gene expression; however, little is known about the development and organization of the mushroom bodies of this insect. The present account bridges this gap by demonstrating that the organization of the Tribolium mushroom bodies is strikingly like that of the fruit fly, with the significant exception that the timeline of neurogenesis is shifted so that the last population of Kenyon cells is born entirely after adult eclosion. Tribolium Kenyon cells are generated by two large neuroblasts per hemisphere and segregate into an early-born delta lobe subpopulation followed by clear homologs of the Drosophila gamma, alpha'/beta' and alpha/beta lobe subpopulations, with the larval-born cohorts undergoing dendritic reorganization during metamorphosis. BrdU labeling and immunohistochemical staining also reveal that a proportion of individual Tribolium have variable numbers of mushroom body neuroblasts. If heritable, this variation represents a unique opportunity for further studies of the genetic control of brain region size through the control of neuroblast number and cell cycle dynamics.

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“…albimanus and Aedes aegypti. piggyBac has also been used to transform a coleopteran species, the red flour beetle, Tribolium castaneum, (Lorenzen et al, 2002) and a hymenopteran species, the sawfly, Athalia rosae, (Sumitani et al, 2003). Despite the piggyBac transposon being originally derived from a lepidopteran insect, this vector has been employed to transform only two lepidopteran species, the pink bollworm, Pectinophora gossypiella, (Peloquin et al, 2000) and the silkworm, Bombyx mori (Tamura et al, 2000).…”

Section: Introductionmentioning

confidence: 99%