Cloning and expression analysis of the Cytosolic NADP-malic Enzyme Gene of Apple Fruit

Abstract

Abstract: 【Objective】The purpose of this study is to isolate a fruit-related gene encoding cytosolic NADP-dependent malic enzyme and reveal the correlation between transcription and malic acid content of fruits.【Method】A full-length cDNA was cloned by RT-PCR and RACE (Rapid Amplification of cDNA Ends) from apple fruits, and its expression was studied by Semi-quantitative RT-PCR analysis，The recombinant prokaryotic expression vector pET30a-ME was constructed and expressed in E.coli BL21.【Result】A fruit-related gene encoding cytosolic NADP-dependent malic enzyme was cloned for the frist time and designated as Mal-ME (GeneBank accession No. DQ280492). The recombinant prokaryotic expression vector pET30a-ME expressed a 66kD fusion protein in E.coli BL21. Semi-quantitative RT-PCR analysis showed that its transcription had negative correlation with malic acid content of fruits at different developmental stages.【Conclusion】cytosolic NADP-dependent malic enzyme gene was successfully isolated from apple fruits. This gene can regulate the accumulation of malic acid in fruits negatively during the development.

CLC Number:

S661.1

Li Ming , Yao YuXin,Liu Zhi and Zhai Heng. Cloning and expression analysis of the Cytosolic NADP-malic Enzyme Gene of Apple Fruit[J]. Chinese Agricultural Science Bulletin, 2007, 23(7): 95-95.

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