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2012 | 59 | 4 |

Tytuł artykułu

Preparation of a polyclonal antibody against hypericin synthase and localization of the enzyme in red-pigmented Hypericum perforatum L. plantlets

Autorzy

Warianty tytułu

Języki publikacji

EN

Abstrakty

EN
 Hypericum perforatum is well known for its antidepressant and anti-inflammatory activities, for which hypericin and its derivatives are indicated to be the most active compounds. Hypericin synthase (Hyp-1) is the only protein proven to catalyze the synthesis of hypericin. In this study, the full-length cDNA of Hyp-1 was chemically synthesized according to the Hyp-1 sequence in GenBank (accession no. AY148090) and then cloned into the plasmid pET22b. Hyp-1 was expressed in Escherichia coli BL21 (DE3) and purified with a Ni-NTA column. The purified protein was used to immunize New Zealand white rabbits, from which an antiserum was purified by protein G affinity chromatography. The polyclonal antibody against Hyp-1 provides a valuable tool for the study of hypericin biosynthesis in H. perforatum. Expression of Hyp-1 and the cellular distribution of hypericin were analyzed in different organs of red-pigmented H. perforatum plantlets. The black glands were not the only site of hypericin accumulation and the results indicated that hypericin might be synthesized in mesophyll cells or in tissues of the root and/or stem and then transported to the glands. This work provides a foundation for further investigation of the regulatory mechanism of hypericin synthesis during the development of H. perforatum.

Słowa kluczowe

Wydawca

-

Rocznik

Tom

59

Numer

4

Opis fizyczny

p.639-645,fig.,ref.

Twórcy

autor
  • School of Life Science and Technology, Tongji University, Shanghai, China
autor
  • School of Life Science and Technology, Tongji University, Shanghai, China
autor
  • School of Life Science and Technology, Tongji University, Shanghai, China
autor
  • State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China

Bibliografia

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  • Curtis JD, Lersten NR (1990) Internal secretory structures in Hypericum (Clusiaceae): H. perforatum L. and H. Balearicum L. New Phytol 114: 571-580.
  • Fernandes H, Konieczna M, Kolodziejczyk R, Bujacz G, Sikorski M, Jaskolski M (2008) Crystallization and preliminary crystallographic studies of Hyp-1, a St. John's wort protein implicated in the biosynthesis of hypericin. Acta Crystallogr., Sect F: Struct Biol Cryst Commun 64: 405-408. 
  • Fields PG, Arnason JT, Fulcher RG (1990) The spectral properties of Hypericum perforatum leaves: the implications for photoactivated defenses. Can J Bot 68: 1166-1170.
  • Fornasiero RB, Bianchi A, Pinetti A (1998) Anatomical and ultrastructural observations in Hypericum perforatum L. leaves. J Herbs, Spices and Med Plants 5: 21-23.
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  • Kosuth J, Katkovcinová Z, Olexová P, Cellárová E (2007) Expression of the hyp-1 gene in early stages of development of Hypericum perforatum L. Plant Cell Rep 26: 211-217.  
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  • Liu CD, Kwan D, Saxton RE, McFadden DW (2000) Hypericin and photodynamic therapy decreases human pancreatic cancer in vitro. J Surg Res 93: 137-143. 
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  • Liu WZ, Hu ZH (1999) The secretory structure of Hypericum perforatum and its relation to hypericin accumulation. Acta Bot Sin 41: 369-372.
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  • Taher MM, Lammering GM, Hershey CM, Valerie KC (2002) Mood-enhancing antidepressant St. John's wort inhibits the activation of human immunodeficiency virus gene expression by ultraviolet light. IUBMB Life 54: 357-364. 
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