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苏格兰松中编码铜锌超氧化物歧化酶的cDNA的特征分析

Characterization of cDNAs encoding CuZn-superoxide dismutases in Scots pine.

作者信息

Karpinski S, Wingsle G, Olsson O, Hällgren J E

机构信息

Department of Forest Genetics and Plant Physiology, Faculty of Forestry, Swedish University of Agricultural Sciences, Umeå.

出版信息

Plant Mol Biol. 1992 Feb;18(3):545-55. doi: 10.1007/BF00040670.

Abstract

A Scots pine (Pinus sylvestris L.) cDNA library was screened with two heterologous cDNA probes (P31 and T10) encoding cytosolic and chloroplastic superoxide dismutases (SOD) from tomato. Several positive clones for cytosolic and chloroplastic superoxide dismutases were isolated, subcloned, mapped and sequenced. One of the cDNA clones (PS3) had a full-length open reading frame of 465 bp corresponding to 154 amino acid residues and showed approximately 85% homology with the amino acid sequences of angiosperm cytosolic SOD counterparts. Another cDNA clone (PST13) was incomplete, but encoded a putative protein with 93% homology to pea and tomato chloroplastic superoxide dismutase. The derived amino acid sequence from both cDNA clones matched the corresponding N-terminal amino acid sequence of the purified mature SOD isozymes. Northern blot hybridizations showed that, cytosolic and chloroplastic CuZn-SOD are expressed at different levels in Scots pine organs. Sequence data and Southern blot hybridization confirm that CuZn-SODs in Scots pine belong to a multigene family. The results are discussed in relation to earlier observations of CuZn-SODs in plants.

摘要

用两个分别编码番茄胞质和叶绿体超氧化物歧化酶(SOD)的异源cDNA探针(P31和T10)筛选了欧洲赤松(Pinus sylvestris L.)的cDNA文库。分离、亚克隆、定位并测序了几个胞质和叶绿体超氧化物歧化酶的阳性克隆。其中一个cDNA克隆(PS3)有一个465 bp的全长开放阅读框,对应154个氨基酸残基,与被子植物胞质SOD对应物的氨基酸序列显示出约85%的同源性。另一个cDNA克隆(PST13)不完整,但编码一种与豌豆和番茄叶绿体超氧化物歧化酶有93%同源性的假定蛋白质。两个cDNA克隆推导的氨基酸序列与纯化的成熟SOD同工酶相应的N端氨基酸序列匹配。Northern印迹杂交表明,胞质和叶绿体CuZn-SOD在欧洲赤松器官中的表达水平不同。序列数据和Southern印迹杂交证实欧洲赤松中的CuZn-SOD属于一个多基因家族。结合植物中CuZn-SOD的早期观察结果对这些结果进行了讨论。

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