TY - JOUR
T1 - Extrachromosomal transposition of the transposable element Minos in embryos of the cricket Gryllus bimaculatus
AU - Zhang, Hongjie
AU - Shinmyo, Yohei
AU - Hirose, Ayumi
AU - Mito, Taro
AU - Inoue, Yoshiko
AU - Ohuchi, Hideyo
AU - Loukeris, Thanasis G.
AU - Eggleston, Paul
AU - Noji, Sumihare
N1 - Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2002
Y1 - 2002
N2 - Effective germline transformation of insects has been shown to depend on the right choice of transposon system and selection marker. In this study the promoter region of a Gryllus cytoplasmic actin (GbA3/4) gene was isolated and characterized, and was used to drive the expression of Minos transposase in embryos of the cricket Gryllus bimaculatus. Active Minos transposase was produced in these embryos as monitored through established transposon excision and interplasmid transposition assays. In contrast, Drosophila melanogaster hsp70 promoter, previously used to express Minos transposase in a number of insect species and insect cell lines, failed to produce any detectable Minos transposase activity, as recorded by using the very sensitive transposon excision assay. In addition, the GbA3/4 promoter was found to drive expression of enhanced green fluorescent protein (eGFP) predominantly in vitellophages of the developing Gryllus eggs when a plasmid carrying a GbA3/4 promoter-eGFP fusion gene was transiently injected into embryos. These results strongly support the use of Minos transposons marked with the GbA3/4 promoter-eGFP for the genetic transformation of this emerging model insect species.
AB - Effective germline transformation of insects has been shown to depend on the right choice of transposon system and selection marker. In this study the promoter region of a Gryllus cytoplasmic actin (GbA3/4) gene was isolated and characterized, and was used to drive the expression of Minos transposase in embryos of the cricket Gryllus bimaculatus. Active Minos transposase was produced in these embryos as monitored through established transposon excision and interplasmid transposition assays. In contrast, Drosophila melanogaster hsp70 promoter, previously used to express Minos transposase in a number of insect species and insect cell lines, failed to produce any detectable Minos transposase activity, as recorded by using the very sensitive transposon excision assay. In addition, the GbA3/4 promoter was found to drive expression of enhanced green fluorescent protein (eGFP) predominantly in vitellophages of the developing Gryllus eggs when a plasmid carrying a GbA3/4 promoter-eGFP fusion gene was transiently injected into embryos. These results strongly support the use of Minos transposons marked with the GbA3/4 promoter-eGFP for the genetic transformation of this emerging model insect species.
KW - Cytoplasmic actin promoter
KW - Embryos
KW - Extrachromosomal transposition
KW - Gryllus bimaculatus
KW - Minos
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U2 - 10.1046/j.1440-169X.2002.00654.x
DO - 10.1046/j.1440-169X.2002.00654.x
M3 - Article
C2 - 12392574
AN - SCOPUS:0036413252
VL - 44
SP - 409
EP - 417
JO - Development Growth and Differentiation
JF - Development Growth and Differentiation
SN - 0012-1592
IS - 5
ER -