荞麦（苦荞麦）种子贮藏蛋白的表达、克隆及免疫学分析

Expression, cloning, and immunological analysis of buckwheat (Fagopyrum esculentum Moench) seed storage proteins.

作者信息

Fujino K, Funatsuki H, Inada M, Shimono Y, Kikuta Y

机构信息

Department of Crop Physiology, Graduate School of Agriculture, Hokkaido University, Sapporo 060-8589, Japan.

出版信息

J Agric Food Chem. 2001 Apr;49(4):1825-9. doi: 10.1021/jf0011485.

DOI:10.1021/jf0011485

PMID:11308332

Abstract

cDNA of buckwheat (Fagopyrum esculentum Moench) was isolated from immature seeds harvested 14 days after pollination. Two genes, designated FA02 and FA18, were found to encode legumin-like proteins and were expressed during seed development. The deduced amino acid sequence of FA02 was identical to the N-terminal amino acid domain of BW24KD, which was believed to be a major buckwheat allergen (Urisu, A.; Kondo, Y.; Morita, Y.; Yagi, E.; Tsuruta, M.; Yasaki, T.; Yamada, K.; Kuzuya, H.; Suzuki, M.; Titani, K.; Kurosawa, K. Isolation and characterization of a major allergen in buckwheat seeds. In Current Advances in Buckwheat Research; Shinshu University Press: Matsumoto, Japan, 1995; pp 965--974). It was predicted that FA02 would be cleaved to generate two separate components, a 41.3 kDa alpha-subunit and a 21 kDa beta-subunit. Antiserum was raised against the deduced FA02 beta-subunit, and immunoblotting of total protein from buckwheat seeds (F. esculentum M. and Fagopyrum tartaricum Gaertn.) revealed that several groups of proteins reacted with the antiserum. Polypeptides in the 23--25 kDa range displayed the greatest reactivity.

摘要

从授粉后14天收获的苦荞未成熟种子中分离出苦荞（Fagopyrum esculentum Moench）的cDNA。发现两个基因，命名为FA02和FA18，编码类豆球蛋白蛋白，并在种子发育过程中表达。FA02推导的氨基酸序列与BW24KD的N端氨基酸结构域相同，BW24KD被认为是苦荞的主要过敏原（Urisu, A.; Kondo, Y.; Morita, Y.; Yagi, E.; Tsuruta, M.; Yasaki, T.; Yamada, K.; Kuzuya, H.; Suzuki, M.; Titani, K.; Kurosawa, K. 苦荞种子中主要过敏原的分离与鉴定。见《苦荞研究的当前进展》；信州大学出版社：日本松本，1995年；第965 - 974页）。据预测，FA02将被切割产生两个独立的组分，一个41.3 kDa的α亚基和一个21 kDa的β亚基。制备了针对推导出来的FA02β亚基的抗血清，对苦荞种子（F. esculentum M.和Fagopyrum tartaricum Gaertn.）的总蛋白进行免疫印迹分析，结果显示几组蛋白与抗血清发生反应。23 - 25 kDa范围内的多肽反应性最强。