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Acta Agron Sin ›› 2014, Vol. 40 ›› Issue (12): 2192-2197.doi: 10.3724/SP.J.1006.2014.02192


Sequence Characteristics and Expression of NAD-malic enzyme in Amaranthus hypochondriacus L.

BAI Yun-Feng1,3,NIE Jiang-Ting2,ZHANG Zhong-Liang1,LI Ping1,ZHANG Wei-Feng1,YAN Jian-Jun1,FENG Rui-Yun1,ZHANG Yao1   

  1. 1 Institute of Crop Science, Shanxi Academy of Agricultural Sciences, Taiyuan 030032, China; 2 College of Bioengineering, Shanxi University, Taiyuan 030006, China; 3 China Key Laboratory of Loess Plateau Crop Gene Resources and Germplasm Creation, Ministry of Agriculture, Taiyuan 030006, China
  • Received:2014-02-25 Revised:2014-09-16 Online:2014-12-12 Published:2014-09-26


The NAD(P)-malic enzyme (NAD(P)-ME) found in many metabolic pathways catalyzes the oxidative decarboxylation of L-malate, which results in producing pyruvate, CO2 and NAD(P)H. In C4 plants, NAD(P)-ME plays a key role in photosynthetic carbon fixation. This study was aimed to characterize the AhNAD-ME in dicotyledonous C4 Amaranthus hypochondriacus by sequence analysis, examine the expression patterns of AhNAD-ME gene in different tissues and different durations of illumination time, and construct a recombinant plasmid pEASY-E1 harboring the AhNAD-ME cDNA and then transform the plasmid into E. coli Transette (DE3) for prokaryotic expression after IPTG induction. The result showed that AhNAD-ME contains all of the motifs required for a complete and functional malic enzyme and is localized specifically to the mitochondrial matrix.Semi-quantitative RT-PCR results showed that AhNAD-ME was constitutively expressed in all examined tissues, with different expression levels, and strongly up-regulated under light in the leaf and stem. Results of SDS-PAGE demonstrated that the specific fusion protein with an expected molecular weight was successfully expressed in E. coli transette (DE3) induced by IPTG

Key words: Amaranthus hypochondriacus, AhNAD-ME, Sequence characteristics, Expression pattern, Prokaryotic expression

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