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一种用于大豆和玉米中谷胱甘肽S-转移酶基因家族综合分析的基因组学方法。

A genomics approach to the comprehensive analysis of the glutathione S-transferase gene family in soybean and maize.

作者信息

McGonigle B, Keeler S J, Lau S M, Koeppe M K, O'Keefe D P

机构信息

Nutrition and Health, E.I. du Pont de Nemours and Company, Experimental Station, P.O. Box 80328, Wilmington, Delaware 19880-0328, USA.

出版信息

Plant Physiol. 2000 Nov;124(3):1105-20. doi: 10.1104/pp.124.3.1105.

Abstract

By BLAST searching a large expressed sequence tag database for glutathione S-transferase (GST) sequences we have identified 25 soybean (Glycine max) and 42 maize (Zea mays) clones and obtained accurate full-length GST sequences. These clones probably represent the majority of members of the GST multigene family in these species. Plant GSTs are divided according to sequence similarity into three categories: types I, II, and III. Among these GSTs only the active site serine, as well as another serine and arginine in or near the "G-site" are conserved throughout. Type III GSTs have four conserved sequence patches mapping to distinct structural features. Expression analysis reveals the distribution of GSTs in different tissues and treatments: Maize GSTI is overall the most highly expressed in maize, whereas the previously unknown GmGST 8 is most abundant in soybean. Using DNA microarray analysis we observed increased expression among the type III GSTs after inducer treatment of maize shoots, with different genes responding to different treatments. Protein activity for a subset of GSTs varied widely with seven substrates, and any GST exhibiting greater than marginal activity with chloro-2,4 dinitrobenzene activity also exhibited significant activity with all other substrates, suggesting broad individual enzyme substrate specificity.

摘要

通过在一个大型表达序列标签数据库中进行BLAST搜索以查找谷胱甘肽S-转移酶(GST)序列,我们鉴定出了25个大豆(Glycine max)克隆和42个玉米(Zea mays)克隆,并获得了准确的全长GST序列。这些克隆可能代表了这些物种中GST多基因家族的大多数成员。植物GST根据序列相似性分为三类:I型、II型和III型。在这些GST中,只有活性位点丝氨酸以及“G位点”内或附近的另一个丝氨酸和精氨酸在整个序列中是保守的。III型GST有四个保守序列片段,对应于不同的结构特征。表达分析揭示了GST在不同组织和处理中的分布:玉米GSTI总体上在玉米中表达最高,而之前未知的GmGST 8在大豆中最为丰富。使用DNA微阵列分析,我们观察到在玉米芽经过诱导剂处理后,III型GST的表达增加,不同基因对不同处理有反应。一部分GST对七种底物的蛋白质活性差异很大,并且任何对氯-2,4-二硝基苯表现出大于边际活性的GST对所有其他底物也表现出显著活性,这表明单个酶具有广泛的底物特异性。

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