PURIFICATION OF FLAVANONE 3-BETA-HYDROXYLASE FROM PETUNIA-HYBRIDA - ANTIBODY PREPARATION AND CHARACTERIZATION OF A CHEMOGENETICALLY DEFINED MUTANT

被引：48

作者：

BRITSCH, L

机构：

[1] Biologisches Institut II der Universität, D-7800 Freiburg

来源：

ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS | 1990年 / 276卷 / 02期

关键词：

D O I：

10.1016/0003-9861(90)90731-D

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Flavanone 3β-hydroxylase from Petunia hybrida has been purified to apparent homogeneity utilizing an improved purification protocol including chromatography of the partially purified enzyme on hydroxyapatite, chromatofocusing, and hydrophobic interaction chromatography on phenyl-Superose. The specificity of mouse and rabbit polyclonal antisera directed to flavanone 3β-hydroxylase was demonstrated by Western blotting and immunotitration with crude extracts from wild-type flowers of P. hybrida and with the purified enzyme. Cross-reactivity was observed with flavanone 3β-hydroxylase from extracts of illuminated parsley cells. A Petunia mutant with white flowers, previously shown to lack 3β-hydroxylase activity and to accumulate flavanone glycosides, showed complete absence of the enzyme protein. © 1990.

引用

页码：348 / 354

页数：7

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