Cloning and Characterization of ATP Synthase CF1 a Gene from Sweet Potato
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Sweet potato (Ipomoea batatas L. Lam) is the seventh important crop in the world and has been planted widely due to its high and stable yield, strong adaptability, rich nutrient content, high photosynthetic capacity, being easy to manage and multiple uses. ATPase is the key enzyme for energy metabolism in the vegetation and occurs from eukaryotic chloroplast, mitochondria to bacteria, and widely takes part in oxidative phosphorylation and photosynthetic phosphorylation.
ATP synthase CF1 α subunit protein is a key enzyme for energy metabolism in plant kingdom, and plays an important role in multiple cell processes. In this study, the complete atpA gene (accession no. JN247444) was cloned from sweet potato (Ipomoea batatas L. Lam) by reverse transcriptasepolymerase chain reaction (RT-PCR). This atpA gene contains an open reading frame (ORF) of 1524 bp coding for a peptide of 507 amino acids with a molecular mass of 55.36 kD.
Sequence analysis showed that atpA gene from sweet potato has high homology with the other plant chloroplast atpA. The transcript levels of the atpA gene in young leaves, mature leaves, stems and tuberous roots were examined by the digital gene expression profiling (DGE), and then confirmed by semi-quantitative RTPCR. The results demonstrate that the highest transcription of atpA gene was found in young leaves, but it was relatively lower in other three tissues. In addition, the atpA gene was successfully expressed in Escherichia coli.
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