Cloning and expression of an alternative oxidase gene from Lycopersicon esculentum.
A full-length cDNA gene (LeAox1au) was isolated from a cDNA library made from ripening fruit of tomato "UC-82B" (Lycopersicon esculentum), after probing with alternative oxidase (AOX) gene fragments, obtained by degenerate primer PCR. Sequence analysis showed that LeAox1au was 1418 bp long and contained a 1077-bp open reading frame encoding a about 40 kD precursor protein which is processed to a mature protein of 32 kD. Southern blot analysis suggested LeAox1au is present as a single copy in the genome of tomato. RT-PCR analysis indicated LeAox1au was expressed in roots, stems, leaves and cotyledons of tomato plants. A recombinant construct containing the open reading frame sequence of the LeAox1au was transformed into Escherichia coli to express the alternative oxidase precursor protein.
Song
Cong-Feng
CF
Key Lab of Monitoring and Management of Plant Diseases and Insects, Ministry of Agriculture, Department of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China. songcf@njau.edu.cn
Borth
Wayne
W
Wang
Jin-Sheng
JS
Hu
John-Sheng
JS
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D018628:Gene Dosage; D018551:Solanum lycopersicum / Q000201:enzymology* / Q000235:genetics; D024101:Mitochondrial Proteins; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000235:genetics*; D010940:Plant Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D000099188:Alternative Oxidase
D024101:Mitochondrial Proteins; D010088:Oxidoreductases; D010940:Plant Proteins; D000099188:Alternative Oxidase