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首页> 外文期刊>Nucleic Acids Research >Cloning and characterization of a cDNA encoding a bacteriophage-type RNA polymerase from the higher plant Chenopodium album.
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Cloning and characterization of a cDNA encoding a bacteriophage-type RNA polymerase from the higher plant Chenopodium album.

机译：从高等植物藜属专辑中克隆和鉴定编码噬菌体型RNA聚合酶的cDNA。

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摘要

A full-length cDNA was cloned from Chenopodium album coding for a single subunit bacteriophage-type RNA polymerase. The cDNA isolated from an actively growing cell suspension culture recognized a 3.8 kb transcript on Northern blots. The open readingframe comprised 987 amino acids with a predicted molecular mass of 112 kDa. A comparison of the protein sequence with those of the two known fungal mitochondrial RNA polymerases, from Saccharomyces cerevisiae and Neurospora crassa, revealed extensive homology between the three enzymes, with complete conservation of all catalytically essential amino acids. The putative mitochondrial RNA polymerase from C.album, as well as homologous sequences from rice and barley, which have been partially cloned, lacked two catalytically non-essential regions of up to 176 amino acids near the C-terminus present in the two fungal mitochondrial RNA polymerases. The extreme N-terminus of the cloned C.album RNA polymerase displayed features of a potential mitochondrial transit sequence. In phylogenetic trees constructed to compare the evolutionary relationships between the different single subunit RNA polymerases, the C.album sequence forms a subgroup together with the S.cerevisiae and the N.crassa mitochondrial RNA polymerases, well separated from both bacteriophage enzymes and plasmid-encoded RNA polymerases found in mitochondria of many fungi and some higher plants.
机译：从Chenopodium专辑克隆全长cDNA，编码单个亚基噬菌体型RNA聚合酶。从活跃生长的细胞悬浮培养物中分离的cDNA在Northern印迹上识别出3.8 kb的转录本。开放阅读框包含987个氨基酸，预测的分子量为112 kDa。将该蛋白质序列与酿酒酵母和芥菜神经孢菌的两种已知真菌线粒体RNA聚合酶的蛋白质序列进行比较，发现这三种酶之间具有广泛的同源性，并且完全保留了所有催化必需氨基酸。假单胞菌的假定线粒体RNA聚合酶以及水稻和大麦的部分同源序列已部分克隆，在两个真菌线粒体的C末端附近缺少两个催化非必需区域，最多不包含176个氨基酸。 RNA聚合酶。克隆的C.album RNA聚合酶的极端N端显示潜在的线粒体转运序列的特征。在用来比较不同的单个亚基RNA聚合酶之间进化关系的系统树中，C.album序列与S.cerevisiae和N.crassa线粒体RNA聚合酶一起形成了一个亚组，与细菌噬菌体酶和质粒编码的碱基相分离RNA聚合酶存在于许多真菌和一些高等植物的线粒体中。

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