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China Journal of Chinese Materia Medica. 2020, (01) Core Bilingual

Cloning and expression analysis of 5-phosphomevalonate kinase gene （CcPMK） in Cinnamomum camphoraChinese Full TextEnglish Full Text (MT)

ZHENG Han;YU Mu-yao;PU Chun-juan;CHEN Mei-lan;LI Fu-quan;SHEN Ye;HUANG Lu-qi;Tianjin University of Traditional Chinese Medicine;State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences;Hulunbeier Mongolian Medical Hospital;

Abstract: The 5-phosphomevalonate kinase（PMK） is a key enzyme in mevalonate（MVA） pathway which reversibly catalyzes the phosphorylation of mevalonate 5-phosphate（MVAP） to form mevalonate-5-diphosphate（MVAPP） in the presence of ATP and divalent metal ion such as Mg2+. In this research, on the basis of the transciptome database of Cinnamomum camphora, the PMK was cloned by cDNA from C. camphora, and was named CcPMK（GenBank number KU886266）. The ORF of CcPMK was composed of 1 545 bp, encoding 514 amino acids. The bioinformatics analysis of CcPMK indicated that the molecular weight of the encoded protein was 56.14 kDa, with a theoretically isoelectric point of 7.64, and there was no signal peptide and transmembrane structure in putative protein. By multiple sequence alignment and phylogenetic tree analysis, we found that similarity between CcPMK and PMK amino acid sequence of other plants was as high as 75%. Among the similar sequences, 45% of them belonged to the alpha helix, while 16% belonged to the beta strand. CcPMK obtained 3 PMK protein family motifs and 1 ATP binding site Gly-Leu-Gly-Ser-Ser-Ala-Ala, and its 3 D structure contained a catalytic pocket structure, proving CcPMK as a member of PMK gene family. The result of phylogenetic tree showed that CcPMK was closely related to monocotyledon plants such as Phonenix dactylifera. The results of the Real-time PCR indicated that the expression level of CcPMK in borneol type was higher than that in linalool type, cineol type, iso-nerolidol type and camphor type. CcPMK expressed highest in roots and lowest in branches. Our results revealed that the expression level of CcPMK was different among five chemical types and different plant tissues, and the research provides foundation for further study of the terpenoids biosynthetic pathway in C. camphora.
Keywords：

Cinnamomum camphora; 5-phosphomevalonate kinase; chemical type; bioinformatics; expression analysis;

  • DOI：

    10.19540/j.cnki.cjcmm.20191104.104

  • Series：

    (E) Medicine ＆ Public Health; (A) Mathematics/ Physics/ Mechanics/ Astronomy; (D) Agriculture

  • Subject：

    Biology; Crop

  • Classification Code：

    Q943.2;S567.19

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