Submitted to: Insect Biochemistry and Molecular Biology
Publication Type: Peer reviewed journal
Publication Acceptance Date: 4/16/2008
Publication Date: 7/15/2008
Citation: Siebert, K.S., Lorenzen, M.D., Brown, S.J., Park, Y., Beeman, R.W. 2008. Tubulin superfamily genes in Tribolium castaneum, and use of a Tubulin promoter to drive transgene expression. Insect Biochemistry and Molecular Biology 38: 749-755. Interpretive Summary: Recent advances in genomics and bioinformatics have opened the door to rapid discovery and analysis of genes that regulate important biological phenomena. Gene transfer technology is needed to fully understand and exploit potentially useful genes discovered through genomics. In this research, we developed new techniques that can be used to understand the regulation of expression of biopesticide target genes, such as those involved in central nervous or molting processes, diuresis, and other vital physiological systems. Such techniques will be used to discover, characterize and exploit insect control genes in pest insects.
Technical Abstract: The use of native promoters to drive transgene expression has facilitated overexpression studies in Drosophila and other insects. We identified twelve 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 a-Tubulin1 (TcaTub1) 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 TcaTub1 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. Nevertheless the TcaTub1 promoter was less active than the Drosophila hsp70 promoter in driving piggyBac transposase in helper/donor hybrids.