Cloning and Sequence Analysis of Rice Mitochondrial Phosphate Transporter Gene by One-step RT-PCR
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    Abstract:

    Gene-specific primers were designed according to the known sequence of mitochondrial phosphate transporter gene (MPT), and the rice MPT (OsMPT) cDNA fragment of 1118 bp was cloned by One-step reverse transcription PCR (RT-PCR) from Cpslo17, a wide-compatibility variety of rice (Oryza saliva). DNA sequencing analysis indicated that it carried an entire open reading frame and encoded a protein of 368 amino acid residues. The deduced amino acid sequence of OsMPT showed similarities of 93.5%, 85.6%, 83.8%, 83.7% and 81. 1% with those from maize, soybean, Lotus japonicus, Betula pendul and A rabidopsis thaliana, respectively. The hydropathy profiles of the deduced amino acid sequence predicted the existence of six membrane-spanning domains, which are highly conserved in the mitochondrial transporter family. The N-terminal amino acids were enriched with arginine, alanine and serine, and a signal peptide targeted in mitochondria was predicted by iPSORT. Further analysis of ecoding segment of OsMPT revealed that it contained 6 exons and 5 introns. The average length of introns and exons were 467 bp and 186.3 bp respectively; and the average GC content of introns and exons are 35.5% and 54.0% respectively. RT-PCR results showed relative high-level expression of OsMPT in leaves of both jopnica and indica rices, so was in both vegetative and reproductive tissues of Cpslo 17. Cloning and expression analysis of OsMPT will provide a base for the researches of its structure and biological function.

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王峰, 黄璐圆, 王宏斌, 刘兵, 王金发.一步法RT—PCR克隆水稻线粒体磷转运蛋白基因及其序列特征分析[J].热带亚热带植物学报,2005,13(4):303~309

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