cDNA cloning and induction of tyrosine hydroxylase gene from the diamondback moth, Plutella xylostella.
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.
Hwang
Se Hui
SH
Department of Applied Biology, College of Agriculture and Life Science, Kangwon National University, Chuncheon, Korea.
Cho
Saeyoull
S
Park
Yong Chul
YC
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D016000:Cluster Analysis; D017124:Conserved Sequence / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004790:Enzyme Induction / Q000502:physiology; D008969:Molecular Sequence Data; D009036:Moths / Q000201:enzymology*; D010766:Phosphorylation; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012694:Serine / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000096:biosynthesis* / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
D017931:DNA Primers; D018076:DNA, Complementary; D012694:Serine; D014446:Tyrosine 3-Monooxygenase