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来自槐树(Sophora japonica)的树皮和种子凝集素的分子克隆。

Molecular cloning of the bark and seed lectins from the Japanese pagoda tree (Sophora japonica).

作者信息

Van Damme E J, Barre A, Rouge P, Peumans W J

机构信息

Laboratory for Phytopathology and Plant Protection, Katholieke Universiteit Leuven, Belgium.

出版信息

Plant Mol Biol. 1997 Feb;33(3):523-36. doi: 10.1023/a:1005781103418.

Abstract

cDNA clones encoding the bark and seed lectins from Sophora japonica were isolated and their sequences analyzed. Screening of a cDNA library constructed from polyA RNA isolated from the bark resulted in the isolation of three different lectin cDNA clones. The first clone encodes the GalNAc-specific bark lectin which was originally described by Hankins et al. [9] whereas the other clones encode the two isoforms of the mannose/glucose-specific lectin reported by Ueno et al. [34]. Molecular cloning of the seed lectin genes revealed that Sophora seeds contain only a GalNAc-specific lectin which is highly homologous to though not identical with the GalNAc-specific lectin from the bark. All lectin polypeptides are translated from mRNAs of ca. 1.3 kb encoding a precursor carrying a signal peptide. In the case of the mannose/glucose-specific bark lectins this precursor is post-translationally processed in two smaller peptides. Alignment of the deduced amino acid sequences of the different clones revealed striking sequence similarities between the mannose/glucose-binding and the GalNAc-specific lectins. Furthermore, there was a high degree of sequence homology with other legume lectins which allowed molecular modelling of the Sophora lectins using the coordinates of the Pisum sativum, Lathyrus ochrus and Erythrina corallodendron lectins.

摘要

分离出了编码日本槐树皮和种子凝集素的cDNA克隆并对其序列进行了分析。从树皮中分离的聚腺苷酸RNA构建的cDNA文库筛选,得到了三个不同的凝集素cDNA克隆。第一个克隆编码最初由汉金斯等人[9]描述的GalNAc特异性树皮凝集素,而其他克隆编码上野等人[34]报道的甘露糖/葡萄糖特异性凝集素的两种同工型。种子凝集素基因的分子克隆表明,日本槐种子只含有一种GalNAc特异性凝集素,它与树皮中的GalNAc特异性凝集素高度同源但不完全相同。所有凝集素多肽均由约1.3kb的mRNA翻译而来,该mRNA编码一个带有信号肽的前体。对于甘露糖/葡萄糖特异性树皮凝集素,该前体在翻译后被加工成两个较小的肽段。不同克隆推导的氨基酸序列比对显示,甘露糖/葡萄糖结合凝集素和GalNAc特异性凝集素之间存在显著的序列相似性。此外,与其他豆科凝集素存在高度的序列同源性,这使得可以利用豌豆、山黧豆和刺桐凝集素的坐标对日本槐凝集素进行分子建模。

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