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Molecular cloning, characterization and expression of atpA and atpB genes from Ginkgo biloba

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Abstract

The chloroplast ATP synthase (ATPase) utilizes the energy of a transmembrane electrochemical proton gradient to drive the synthesis of ATP from ADP and phosphate. The chloroplast ATPase α and β subunits are the essential components of multisubunit protein complex. In this paper, the full-length cDNA and genomic DNA of ATPase α (designated as GbatpA) and β (designated as GbatpB) subunit genes were isolated from Ginkgo biloba. The GbatpA and GbatpB genes were both intronless. The coding regions of GbatpA and GbatpB were 1530 bp and 1497 bp long, respectively, and their deduced amino acid sequences showed high degrees of identity to those of other plant ATPase α and β proteins, respectively. The expression analysis by RT-PCR revealed that GbatpA and GbatpB both expressed in tissue-specific manners in G. biloba and might involve in leaf development. The recombinant GbATPB protein was successfully expressed in E. coli strain using pET28a vector with ATPase activity as three times high as the control, and the results showed that the molecular weight of the recombinant protein was about 54 kDa, a size that was in agreement with that predicted by bioinformatics analysis. This study provides useful information for further studying on overall structure, function and regulation of the chloroplast ATPase in G. biloba, the so-called “living fossil” plant as one of the oldest gymnosperm species.

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Abbreviations

ATPase:

ATP synthase

CF0 :

coupling factor 0

CF1 :

coupling factor 1

H+-ATPase:

proton-translocating ATPase

IPTG:

isopropyl-β-D-thiogalactoside

PSI:

photosystem I

PSII:

photosystem II

RACE:

rapid amplification of cDNA ends

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Authors and Affiliations

  1. College of Horticulture and Gardening, Yangtze University, Jingzhou, 434025, People’s Republic of China

    Feng Xu & Rong Cai

  2. College of Horticulture Science and Engineering, Shandong Agricultural University, Tai’an, 271018, Peope’s Republic of China

    Feng Xu, Liu-ji Chen & Shu-han Cheng

  3. College of Life Science and Engineering, Huanggang Normal University, Huangang, 438000, People’s Republic of China

    Shui-yuan Cheng

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  1. Feng Xu
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  2. Rong Cai
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  3. Shui-yuan Cheng
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  4. Liu-ji Chen
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  5. Shu-han Cheng
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Correspondence to Shui-yuan Cheng.

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These authors contributed equally to this work

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Xu, F., Cai, R., Cheng, Sy. et al. Molecular cloning, characterization and expression of atpA and atpB genes from Ginkgo biloba . Biologia 63, 526–534 (2008). https://doi.org/10.2478/s11756-008-0093-0

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