IDENTIFICATION, CHARACTERIZATION, AND ANALYSIS OF CDNA AND GENOMIC SEQUENCES ENCODING 2 DIFFERENT SMALL HEAT-SHOCK PROTEINS IN HORDEUM-VULGARE

被引:21
作者
MARMIROLI, N
PAVESI, A
DICOLA, G
HARTINGS, H
RAHO, G
CONTE, MR
PERROTTA, C
机构
[1] EXPTL INST CEREAL RES,I-24100 BERGAMO,ITALY
[2] UNIV LECCE,DEPT BIOL,GENET LAB,I-73100 LECCE,ITALY
关键词
BARLEY; CDNA; GENOMIC CLONE; HEAT SHOCK; NUCLEOTIDE SEQUENCE; SMALL HEAT SHOCK PROTEINS;
D O I
10.1139/g93-148
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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.
引用
收藏
页码:1111 / 1118
页数:8
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