caa a22 4500 477101119 CHVBK 20180405111541.0 cr unu---uuuuu 170330e19960201xx s 000 0 eng 10.1007/BF00206244 doi (NATIONALLICENCE)springer-10.1007/BF00206244 The alternative oxidase is encoded in a multigene family in soybean [Elektronische Daten] [James Whelan, A. Millar, David Day] The copy number of the alternative oxidase gene, Aox, was investigated in soybean (Glycine max L.) using a Polymerase chain reaction (PCR) approach to amplify fragments from soybean genomic DNA. The primers used were based on absolutely conserved regions of Aox cDNA clones from a variety of plant species and the yeast Hansenula anomala. After subcloning of the 170-bp PCR products, 12 individual colonies were sequenced. Eleven plasmids yielded inserts representing three sequences in the ratio 4∶3∶4 (Aox1-3). The sequence of Aox1 was 100% identical at the nucleic acid level to the published full-length cDNA from soybean. The other two sequences were 60-75% identical to Aox1 and to each other at the nucleic acid and protein levels. Similar analysis of Nicotiana tabacum L. revealed an additional gene copy with high homology to the soybean Aox2 sequence. Genomic DNA from soybean cut with Hind III and probed with the full-length Aox1 yielded a single positive band of 6.5 kb; when the same genomic blot was probed with a mixture of all three PCR fragments, three bands of 9 kb, 6.5 kb and 3 kb were detected. Reverse transcription-PCR performed on total RNA from various soybean tissues, followed by hybridisation with the three Aox sequences individually, revealed differential expression of the Aox genes between cotyledons and leaves. It is suggested that soybean contains a multigene Aox family. The implication of this for the understanding of alternative oxidase expression and regulation in plant tissues is discussed. Springer-Verlag, 1996 Alternative oxidase nationallicence Gene family nationallicence Glycine nationallicence Mitochondrion nationallicence AOX ( Aox ) : alternative oxidase protein (gene) nationallicence bp : base pairs nationallicence PCR : polymerase chain reaction nationallicence RT-PCR : reverse transcription-PCR nationallicence Whelan James Division of Biochemistry and Molecular Biology, Faculty of Science, and The Cooperative Research Centre for Plant Science, Australian National University, 0200, Canberra, ACT, Australia aut Millar A. Division of Biochemistry and Molecular Biology, Faculty of Science, and The Cooperative Research Centre for Plant Science, Australian National University, 0200, Canberra, ACT, Australia aut Day David Division of Biochemistry and Molecular Biology, Faculty of Science, and The Cooperative Research Centre for Plant Science, Australian National University, 0200, Canberra, ACT, Australia aut Planta Springer-Verlag 198/2(1996-02-01), 197-201 0032-0935 198:2<197 1996 198 425 https://doi.org/10.1007/BF00206244 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00206244 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Whelan James Division of Biochemistry and Molecular Biology, Faculty of Science, and The Cooperative Research Centre for Plant Science, Australian National University, 0200, Canberra, ACT, Australia aut NATIONALLICENCE 700 1- Millar A. Division of Biochemistry and Molecular Biology, Faculty of Science, and The Cooperative Research Centre for Plant Science, Australian National University, 0200, Canberra, ACT, Australia aut NATIONALLICENCE 700 1- Day David Division of Biochemistry and Molecular Biology, Faculty of Science, and The Cooperative Research Centre for Plant Science, Australian National University, 0200, Canberra, ACT, Australia aut NATIONALLICENCE 773 0- Planta Springer-Verlag 198/2(1996-02-01), 197-201 0032-0935 198:2<197 1996 198 425 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer