Department of Applied Biology, College of Agriculture and Life Science, Kangwon National University, Chuncheon, Korea.
Arch Insect Biochem Physiol. 2010 Oct;75(2):107-20. doi: 10.1002/arch.20384.
We cloned a full-length tyrosine hydroxylase cDNA from the integument of the diamondback moth, Plutella xylostella. In the phylogenetic tree, tyrosine hydroxylase (PxTH) clustered with the other lepidopteran THs. Serine residues in the PxTH sequence, namely Ser(24), Ser(31), Ser(35), Ser(53), and Ser(65), were predicted to be the target sites for phosphorylation based on PROSITE analysis. In particular, Ser(35) of PxTH is highly conserved across a broad phylogenetic range of animal taxa including rat and human. Western blot analysis using both PxTH-Ab1 and PxTH-Ab2 polyclonal antibodies verified the expression of PxTH in all life cycle stages of P. xylostella, namely the larval, pupal, and adult stages. To examine the possible immune function of PxTH in P. xylostella, PxTH gene expression was investigated by RT-PCR and western blotting analysis after challenging P. xylostella with bacteria. PxTH expression was elevated 1 h post-infection and was continued till 12 h of post-infection relative to control larvae injected with sterile water.
我们从小菜蛾的表皮中克隆了全长的酪氨酸羟化酶 cDNA。在系统发育树中，酪氨酸羟化酶（PxTH）与其他鳞翅目 TH 聚类在一起。PxTH 序列中的丝氨酸残基，即 Ser(24)、Ser(31)、Ser(35)、Ser(53)和 Ser(65)，根据 PROSITE 分析预测是磷酸化的靶位点。特别是 PxTH 的 Ser(35)在包括大鼠和人类在内的广泛动物分类群中高度保守。使用 PxTH-Ab1 和 PxTH-Ab2 多克隆抗体进行的 Western blot 分析验证了 PxTH 在小菜蛾所有生命周期阶段的表达，即幼虫、蛹和成虫阶段。为了研究 PxTH 在小菜蛾中的可能免疫功能，通过 RT-PCR 和 Western blot 分析在细菌攻击小菜蛾后检测了 PxTH 基因的表达。与用无菌水注射的对照幼虫相比，感染后 1 小时 PxTH 表达升高，并持续到感染后 12 小时。
