Cloning and Expression Analysis of CCoAOMT Gene from Dendrocalamus sinicus

CHEN Ling-na,GUO Xiao-juan and YANG Han-qi

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Journal of Plant Genetic Resources ›› 2019, Vol. 20 ›› Issue (2) : 476-483. DOI: 10.13430/j.cnki.jpgr.20180731002
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Cloning and Expression Analysis of CCoAOMT Gene from Dendrocalamus sinicus

  • CHEN Ling-na,GUO Xiao-juan and YANG Han-qi
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Abstract

Caffeoyl-CoA O-methyltransferase (CCoAOMT) is one of the key enzymes in lignin biosynthesis. By reversible transcription PCR (RT-PCR) technique, a CCoAOMT gene (DsCCoAOMT) was cloned from the bamboo shoot of Dendrocalamus sinicus. The full-length cDNA of DsCCoAOMT was 777 base pairs that encoded a protein of 238 amino acids with predicted molecular mass of 28.85kDa and a basic isoelectric point of 5.35. The deduced protein was rich in helix and coin. The phylogenetic analysis revealed the highest sequence homology of DsCCoAOMT in relative to CCoAOMT from the poaceous plants, followed by the monocotyledon of non-graminaceae and gymnosperms and then some dicotyledons. That suggested a differentiation of CCoAOMT between monocotyledons and dicotyledons before angiosperms evolution. According to the analysis of codon bias, the DsCCoAOMT had high bias toward the synonymous codons with G and C at third codon position. The results of codon usage frequency showed that heterologous expression of DsCCoAOMT might be applicable in the microorganism yeast expression system and in plants Nicotiana tabacum and Solanum lycopersicum. By real-time quantitative PCR, the DsCCoAOMT gene was highly expressed in the shoots, followed by the culms and the leaves at last. As the shoots developed, DsCCoAOMT is up-regulated at the higher level, suggesting that the gene might play an important regulation role in fast-growing D. sinicus. Thus, this study might be helpful for further understanding the function and molecular mechanism of the DsCCoAOMT gene.

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Dendrocalamus sinicus / CCoAOMT gene / gene clone / bioinformatics analysis / gene expression

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CHEN Ling-na,GUO Xiao-juan and YANG Han-qi. Cloning and Expression Analysis of CCoAOMT Gene from Dendrocalamus sinicus. Journal of Plant Genetic Resources. 2019, 20(2): 476-483 https://doi.org/10.13430/j.cnki.jpgr.20180731002

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Funding

the Fundamental Research Funds for the Central Non-profit Research Institution of CAF
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