Molecular cloning and characterization of a recombinant Bombyx mori tyramine-β-hydroxylase in a silkworm cell line using a baculovirus expression vector system

Ahmed M.H. Ali, Nazmul Hasan, Renkai Guo, Hiroto Ohta, Akinori Hirashima

研究成果: ジャーナルへの寄稿学術誌査読

4 被引用数 (Scopus)

抄録

Octopamine (OA) and tyramine (TA) are biogenic amines that act as neurotransmitters, neurohormones, and neuromodulators in the invertebrate nervous system. Tyramine-β-hydroxylase (TβH) catalyzes the biosynthesis of OA from TA. In this study, cDNA encoding Bombyx mori TβH (BmTβH) was cloned from the brain of the silkworm B. mori. The BmTβH mRNA comprised 2204 nucleotide residues and contained an open reading frame encoding 592 amino acids. The deduced amino acid sequence shared homology to several proteins belonging to the insect TβH family. Functional expression of the cloned cDNA was obtained using a B. mori baculovirus expression vector system. Western blot analysis revealed an immunoreactive band with a molecular mass of ~. 67.4. kDa. Reverse-phase high-performance liquid chromatography (HPLC) was used to identify the products formed during incubation of the enzyme reaction mixture. The optimum pH and temperature for the conversion of TA to OA were 7.5 and 25. °C, respectively. During incubation, the reaction was linear for the first 30. min at 25. °C and pH. 7.5. Inhibitory experiments carried out with various concentrations of an inhibitor showed that this method can be used for screening of BmTβH inhibitors.

本文言語英語
ページ(範囲)221-227
ページ数7
ジャーナルJournal of Asia-Pacific Entomology
17
3
DOI
出版ステータス出版済み - 9月 2014

!!!All Science Journal Classification (ASJC) codes

  • 昆虫科学

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