Transgene expression from the Tribolium castaneum Polyubiquitin promoter
Transgene expression from the Tribolium castaneum Polyubiquitin promoter
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DOI：
10.1046/j.1365-2583.2002.00349.x
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发表时间：
2002-10-01
影响因子：
2.6
通讯作者：
Beeman, RW
中科院分区：
文献类型：
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作者：
Lorenzen, MD;Brown, SJ;Beeman, RW
在许多物种中，高度保守的泛素蛋白表达于具有强启动子的基因，这使得这些启动子在驱动转基因表达方面具有吸引力。本文报道了赤霉菌多聚泛素(TcPUb)基因的克隆和鉴定。我们将TcPUb启动子放在卡氏锥虫眼色基因TC朱砂(TCV)编码区的上游，并将此构建物注射到TCV缺失菌株的胚胎中。在胚胎发生过程中瞬时表达TCV可以完全挽救突变的幼虫表型。然后，我们将TcPUb-TCV嵌合体整合到了猪Bac供体中。通过挽救眼睛色素沉着，很容易识别得到的胚系转化子，说明了TcPUb启动子用于驱动转基因表达的潜力。
The highly conserved Ubiquitin proteins are expressed from genes with strong, constitutively active promoters in many species, making these promoters attractive candidates for use in driving transgene expression. Here we report the cloning and characterization of the Tribolium castaneum Polyubiquitin (TcPUb ) gene. We placed the TcPUb promoter upstream of the coding region of the T. castaneum eye-colour gene Tc vermilion (Tcv ) and injected this construct into embryos from a Tcv -deficient strain. Transient expression of Tcv during embryogenesis resulted in complete rescue of the larval mutant phenotype. We then incorporated the TcPUb-Tcv chimera into a piggyBac donor. Resulting germline transformants were easily recognized by rescue of eye pigmentation, illustrating the potential of the TcPUb promoter for use in driving transgene expression.