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cDNA cloning and induction of tyrosine hydroxylase gene from the diamondback moth,Plutella xylostella
Authors:Se Hui Hwang  Saeyoull Cho  Yong Chul Park
Affiliation:1. Department of Applied Biology, College of Agriculture and Life Science, Kangwon National University, Chuncheon, Korea;2. Agriculture and Life Sciences Research Institute, Kangwon National University, Chuncheon, Korea;3. Department of Plant Biotechnology Program, Division of Biotechnology, School of Biotechnology, Kangwon National University, Chuncheon, Korea
Abstract: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 Ser24, Ser31, Ser35, Ser53, and Ser65, were predicted to be the target sites for phosphorylation based on PROSITE analysis. In particular, Ser35 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. © 2010 Wiley Periodicals, Inc.
Keywords:Plutella xylostella  tyrosine hydroxylase  phylogenetic tree  PROSITE analysis  immune
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