MOLECULAR-CLONING OF COMPLEMENTARY-DNA ENCODING THE LIGNIN-FORMING PEROXIDASE FROM TOBACCO - MOLECULAR ANALYSIS AND TISSUE-SPECIFIC EXPRESSION
MOLECULAR-CLONING OF COMPLEMENTARY-DNA ENCODING THE LIGNIN-FORMING PEROXIDASE FROM TOBACCO - MOLECULAR ANALYSIS AND TISSUE-SPECIFIC EXPRESSION
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DOI：
10.1073/pnas.84.21.7542
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发表时间：
1987-11-01
影响因子：
11.1
通讯作者：
ROTHSTEIN, S
中科院分区：
文献类型：
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作者：
LAGRIMINI, LM;BURKHART, W;ROTHSTEIN, S
植物过氧化物酶在木质素形成和伤口愈合中起主要作用，并且被认为参与生长素催化和对病原体攻击的防御。阴离子过氧化物酶同工酶的功能在烟草中得到了最好的理解。这些同工酶催化木质素聚合物的形成，并在次生植物细胞壁中的木质素、纤维素和伸展蛋白之间形成刚性交联。本文报道了烟草阴离子过氧化物酶同工酶的纯化及其部分氨基酸序列。从氨基酸序列推导出的寡核苷酸探针用于筛选烟草叶片cDNA文库，并分离出1200个碱基对的cDNA克隆，并对其进行完整测序。预测的氨基酸序列显示一个22个氨基酸的信号肽和一个302个氨基酸的成熟蛋白（Mr，332，311）。的氨基酸序列进行了比较，从辣根和芜菁的阳离子过氧化物酶，并被发现是52%和46%的同源性，分别。通过RNA印迹分析，发现烟草同工酶的信使在茎组织中丰富，而在叶和根组织中表达水平很低。在基因组DNA印迹上发现了该基因的四个可区分的拷贝。基因拷贝数可能反映了烟草异源四倍体的性质。
Plant peroxidases play a major role in lignin formation and wound healing and are believed to be involved in auxin catabolism and defense to pathogen attack. The function of the anionic peroxidase isozymes is best understood in tobacco. These isozymes catalyze the formation of the lignin polymer and form rigid cross-links between lignin, cellulose, and extensin in the secondary plant cell wall. We report the purification of the anionic peroxidase isozymes from tobacco and their partial amino acid sequence. An oligonucleotide probe deduced from the amino acid sequence was used to screen a tobacco leaf cDNA library and a 1200-base-pair cDNA clone was isolated and sequenced in its entirety. The predicted amino acid sequence revealed a 22-amino acid signal peptide and a 302-amino acid mature protein (Mr, 332,311). The amino acid sequence was compared to that of the cationic peroxidases from horseradish and turnip and was found to be 52% and 46% homologous, respectively. By RNA blot analysis, the messenger for the tobacco isozyme was found to be abundant in stem tissue while expressed at very low levels in leaf and root tissue. Four distinguishable copies of the gene were found on genomic DNA blots. The gene copy number may reflect the allotetraploid nature of Nicotiana tabacum.