Siebert Kendra S, Lorenzen Marcé D, Brown Susan J, Park Yoonseong, Beeman Richard W
Department of Entomology, Kansas State University, Manhattan, KS 66506, USA.
Insect Biochem Mol Biol. 2008 Aug;38(8):749-55. doi: 10.1016/j.ibmb.2008.04.007. Epub 2008 May 16.
The use of native promoters to drive transgene expression has facilitated overexpression studies in Drosophila and other insects. We identified 12 Tubulin family members from the genome sequence of the red flour beetle, Tribolium castaneum, and used the promoter from one of these to drive constitutive expression of a transgene. The activity of the T. castaneum alpha-Tubulin1 (TcalphaTub1) putative promoter was pre-tested in conjunction with an eye-color gene, T. castaneum vermilion (Tcv), by transient expression in Tcv-deficient embryos. Such embryos showed complete rescue of larval eyespot pigmentation. We also examined the TcalphaTub1 expression pattern in germline transformants using the enhanced green fluorescent protein (EGFP) reporter. Beetles transformed with this piggyBac-based reporter ubiquitously expressed EGFP at all stages.
使用天然启动子驱动转基因表达促进了在果蝇和其他昆虫中的过表达研究。我们从赤拟谷盗（Tribolium castaneum）的基因组序列中鉴定出12个微管蛋白家族成员，并使用其中一个的启动子驱动转基因的组成型表达。通过在缺乏Tcv的胚胎中瞬时表达，将赤拟谷盗α-微管蛋白1（TcalphaTub1）推定启动子的活性与眼色基因赤拟谷盗朱红色（Tcv）一起进行了预测试。此类胚胎显示幼虫眼点色素沉着完全恢复。我们还使用增强型绿色荧光蛋白（EGFP）报告基因检查了种系转化体中的TcalphaTub1表达模式。用这种基于piggyBac的报告基因转化的甲虫在所有阶段均普遍表达EGFP。
