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Apple ripening-related cDNA clone pAP4 confers ethylene-forming ability in transformed Saccharomyces cerevisiae.

Wilson, I. D.; Zhu, Y.; Burmeister, D. M.; Dilley, D. R.;

Plant Physiology Year: 1993 Vol: 102 Issue: 3 Pages: 783-788 Ref: 30 ref.

1993

บทคัดย่อ

Apple ripening-related cDNA clone pAP4 confers ethylene-forming ability in transformed Saccharomyces cerevisiae.

The apple ripening-related cDNA insert of clone pAP4 was previously shown to have considerable nucleic acid and predicted amino acid sequence similarity to the insert of a tomato ripening-related cDNA clone (pTOM13) known to encode the enzyme ACC oxidase. The cDNA insert from the clone pAP4 was fused between the galactose-inducible promoter and the terminator of the yeast expression vector pYES2. Transformation of Saccharomyces cerevisiae strain F808- with this DNA construct and incubation of the yeast in the presence of D[+]-galactose allowed these cells to convert ACC to ethylene. The transformed yeast converted 1-amino-2-ethylcyclopropane-1-carboxylate isomers to 1-butene with the same 1R,2S-stereoselectivity as achieved by the native ACC oxidase from apples. Both ascorbate and Fe2+ ions stimulated the rate of the production of ethylene from ACC by the transformed yeast, whereas Cu2+ and Co2+ were strongly inhibitory. Northern analysis of the total RNA from non-transformed and transformed yea

st showed that the ability to convert the ACC to ethylene was correlated with the synthesis and accumulation of a novel 1.2 kb mRNA that hybridized to the cDNA clone pAP4. It was concluded that the cDNA sequence of the clone pAP4 encodes ACC oxidase.