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cDNA cloning and induction of tyrosine hydroxylase gene from the diamondback moth, Plutella xylostella
Author(s) -
Hwang Se Hui,
Cho Saeyoull,
Park Yong Chul
Publication year - 2010
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/arch.20384
Subject(s) - plutella , diamondback moth , biology , complementary dna , tyrosine hydroxylase , cloning (programming) , gene , genetics , lepidoptera genitalia , botany , enzyme , biochemistry , computer science , programming language
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. © 2010 Wiley Periodicals, Inc.