Cloning and Expression Analysis of 5-Phosphomevalonate Kinase PfPMK Gene in Perilla frutescens L.

doi:10.7668/hbnxb.20192609

Abstract

Abstract:

In order to investigate the function of 5-phosphomevalonate kinase PMK gene in biosynthesis pathway of terpenoids in Perilla frutescens L.,we analyzed the transcriptome data of Perilla frutescens and mined the reference sequence of PMK.The PMK gene was cloned from Perilla frutescens by gene cloning technique,and was analysed using bioinformatics and Real-time PCR(qRT-PCR).The results showed that the ORF of PfPMK gene was 1 524 bp,encoding 507 amino acids.Bioinformatics analysis showed that the molecular weight of PfPMK was 54.73 ku and the isoelectric point was 5.20,which was a hydrophilic protein.The amino acids of PfPMK were higher homologous with SmPMK,SsPMK,SbPMK and PvPMK,which indicated PfPMK protein was highly conserved in the evolutionary process.The phylogenetic analysis showed that PfPMK was closely related to PMK of Salvia miltiorrhiza and Salvia splendens.WoLF-PSORT predicted that PfPMK protein might be located in the plasma membrane or endoplasmic reticulum.Real-time fluorescence quantitative PCR results showed that PfPMK gene was expressed in roots,stems and leaves of Perilla frutescens,and the expression level in roots was higher than that in leaves and stems.The fluorescence quantitative PCR analysis showed that the expression level of PfPMK gene was higher in the middle and late September.The full length of PfPMK gene was first cloned from Perilla frutescens.Bioinformatics analysis showed that PfPMK gene belonged to 5-phosphomevalonate kinase gene and participates in the biosynthesis of terpenoids in Perilla frutescens.

Key words: Perilla frutescens L., PfPMK, Terpenoids, Bioinformatics, Functional analysis

LI Hui, WEN Chunxiu, LIU Lingdi, WEN Saiqun, TANG Yinghong, JIANG Tao. Cloning and Expression Analysis of 5-Phosphomevalonate Kinase PfPMK Gene in Perilla frutescens L.[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(2): 49-55. doi: 10.7668/hbnxb.20192609.

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Figures/Tables 5

Fig.1 Cloning of PfPMK gene and amino acid sequence

Fig.2 Bioinformatics analysis of PfPMK gene

Fig.3 Protein analysis of PfPMK

Fig.4 Amino acid structure prediction of PfPMK

Fig.5 Expression analysis of PfPMK gene
Different lowercase letters indicate significant difference compared with the control(P<0.05).

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