Marmiroli N, Pavesi A, Di Cola G, Hartings H, Raho G, Conte M R, Perrotta C
Department of Evolutionary Biology, University of Parma, Italy.
Genome. 1993 Dec;36(6):1111-8. doi: 10.1139/g93-148.
In vitro translation of mRNAs prepared from barley (Hordeum vulgare) seedlings (cv. Onice) exposed at 40 degrees C directed the synthesis of major heat shock proteins (HSPs) with molecular masses of 80-90, 70, 42 and 16-22 kDa. A cDNA library prepared from the 40 degrees C mRNAs and screened by differential hybridization led to the isolation of heat shock specific sequences. One of these (Hv hsp18) was confirmed by hybrid-arrested and hybrid-released translation as encoding for an 18-kDa HSP. The barley hsp18 sequence has an open reading frame encoding a 160 amino acid residue 18-kDa protein that is 63% identical to wheat 16.9-kDa HSP (clone C5-8), 54% identical to soybean (Glycine max) 17.5-kDa HSP, and 49% identical to Arabidopsis thaliana 17.6-kDa HSP. Lower similarities were found with class II plant small HSPs such as soybean 17.9-kDa HSP (27%), Pisum sativum 17.7-kDa HSP (30%), wheat (Triticum aestivum) 17.3-kDa HSP (clone Ta hsp 17.3) (30%), and with animal small HSPs and alpha-crystallins. The Hv hsp18 sequence was used to pick up Hv hsp17 genomic sequence encoding for another class I 17-kDa HSP. By computer analysis of the nucleotide sequence the TATA box, two heat shock promoter elements, a metal-ion response element, and the polyadenylation signals were identified. Barley HSP18 has an additional cysteine-rich region when compared with HSP17 mapping at the carboxy terminal end.
从40摄氏度处理的大麦（Hordeum vulgare）幼苗（品种Onice）中制备的mRNA进行体外翻译，可指导合成分子量为80 - 90 kDa、70 kDa、42 kDa和16 - 22 kDa的主要热休克蛋白（HSP）。从40摄氏度处理的mRNA制备cDNA文库，并通过差异杂交筛选，从而分离出热休克特异性序列。其中一个序列（Hv hsp18）经杂交抑制翻译和杂交释放翻译证实编码一种18 kDa的HSP。大麦hsp18序列有一个开放阅读框，编码一个含160个氨基酸残基的18 kDa蛋白，该蛋白与小麦16.9 kDa HSP（克隆C5 - 8）的同源性为63%，与大豆（Glycine max）17.5 kDa HSP的同源性为54%，与拟南芥17.6 kDa HSP的同源性为49%。与II类植物小HSP（如大豆17.9 kDa HSP，同源性27%；豌豆17.7 kDa HSP，同源性30%；小麦（Triticum aestivum）17.3 kDa HSP（克隆Ta hsp 1７.3），同源性30%）以及动物小HSP和α - 晶状体蛋白的相似性较低。Hv hsp18序列用于获取编码另一种I类17 kDa HSP的Hv hsp17基因组序列。通过对核苷酸序列的计算机分析，鉴定出TATA框、两个热休克启动子元件、一个金属离子反应元件和多聚腺苷酸化信号。与定位在羧基末端的HSP17相比，大麦HSP18有一个额外的富含半胱氨酸区域。
