Cloning and expression of an alternative oxidase gene from Lycopersicon esculentum.
Congfeng Song, Wayne B. Borth, Jinsheng Wang, John-Sheng Hu
Abstract
Congfeng Song, Wayne B. Borth, Jinsheng Wang, John-Sheng Hu
Abstract
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.
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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.
Key concepts: Lycopersicon, Open reading frame, Complementary DNA, Biology, Southern blot, Molecular biology, Gene, cDNA library