Robison Mary M, Wolyn David J
Department of Plant Agriculture, Bovey Bldg, University of Guelph, N1G 2W1 Guelph, ON, Canada.
Mol Genet Genomics. 2006 Jan;275(1):68-73. doi: 10.1007/s00438-005-0061-0. Epub 2005 Dec 15.
An inverted repeat in the mitochondrial DNA of carrot mitochondrial type (mitotype) SW3 has been sequenced in the context of its up- and downstream genomic environments. The 3,159 bp repeat encodes rps7 and cox1, but the cox1 reading frame does not encode a stop codon at a position predicted by alignment with other plant cox1 genes; cox1 extends into both of the downstream flanking sequences as unique C-terminal extensions of 224 (cox1-1) or 284 (cox1-2) amino acids. Northern and RT-PCR analyses revealed that rps7 and cox1 are co-transcribed and RNA-edited. In three additional mitotypes of carrot, rps7 and cox1 are co-transcribed, but are present as single copies within the genome. In mitotype FG17, the cox1 gene has a C-terminal extension like cox1-1, but in mitotypes FW2 and FW4, the cox1 genes lack C-terminal extensions and have stop codons and 3' UTRs like those of other plant cox1 genes. The C-terminal extensions are transcribed in SW3 and FG17 and result in longer primary transcripts than those from FW2 and FW4. Western immunoblotting of mitochondrial proteins from all mitotypes detected COX1 products of the predicted unextended size in all mitotypes, irrespective of the presence of a C-terminal extension in the reading frame.
已在胡萝卜线粒体类型(mitotype)SW3的线粒体DNA中的一个反向重复序列及其上下游基因组环境的背景下对其进行了测序。这个3159bp的重复序列编码rps7和cox1,但cox1阅读框在与其他植物cox1基因比对预测的位置上没有编码终止密码子;cox1延伸到两个下游侧翼序列中,分别作为224个(cox1-1)或284个(cox1-2)氨基酸的独特C末端延伸。Northern印迹和RT-PCR分析表明,rps7和cox1是共转录且经过RNA编辑的。在胡萝卜的另外三种线粒体类型中,rps7和cox1是共转录的,但在基因组中以单拷贝形式存在。在mitotype FG17中,cox1基因有一个类似于cox1-1的C末端延伸,但在mitotypes FW2和FW4中,cox1基因没有C末端延伸,并且有像其他植物cox1基因那样的终止密码子和3'UTR。C末端延伸在SW3和FG17中被转录,导致其初级转录本比FW2和FW4的更长。对所有线粒体类型的线粒体蛋白进行的Western免疫印迹检测到,所有线粒体类型中都有预测的未延伸大小的COX1产物,无论阅读框中是否存在C末端延伸。