苹果果实中细胞质型苹果酸酶基因
（NADP-ME）的克隆与表达分析

摘要/Abstract

摘要： 【研究目的】从苹果果实中克隆细胞质型NADP-苹果酸酶（NADP-ME）基因，并且从基因转录水平上研究该基因与苹果果实酸度的关系。【方法】利用RT-PCR和RACE技术克隆NADP-ME基因，半定量RT-PCR分析该基因在高酸和低酸个体不同发育时期果实中的转录情况，并进行原核表达分析。【结果】成功克隆了细胞质型NADP-ME基因的cDNA全长序列，命名为Mal-ME (GenBank注册号：DQ280492)；该基因表达一个约66kD的蛋白；半定量RT-PCR分析表明：不同发育时期的果实中Mal-ME的转录与苹果酸含量呈负相关，即随着Mal-ME转录水平的增强果实含酸量下降。【结论】成功从苹果果实中克隆了细胞质型Mal-ME基因，Mal-ME在果实发育过程中对苹果酸的积累起负调控作用。

关键词: 植被破坏, 植被破坏, 团聚体稳定性, 土壤有机碳

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.

中图分类号:

S661.1

李明,姚玉新,刘志,翟衡. 苹果果实中细胞质型苹果酸酶基因（NADP-ME）的克隆与表达分析[J]. 中国农学通报, 2007, 23(7): 95-95.

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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