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[Cloning and expression analysis of CpPMM gene in Codonopsis pilosula].

Cao Shou-Bo¹, Wang Xiao-Lin¹, J I Jiao-Jiao¹

¹College of Pharmacy, Shanxi Medical University Jinzhong 030600, China.

Zhongguo Zhong Yao Za Zhi = Zhongguo Zhongyao Zazhi = China Journal of Chinese Materia Medica

|November 9, 2020

Summary

The cloned Codonopsis pilosula phosphomannomutase (CpPMM) gene shows high correlation with polysaccharide synthesis. Its expression increases with plant growth, peaking during the fruiting period, mirroring rising polysaccharide content in this medicinal plant.

Keywords:

Codonopsis pilosula CpPMM bioinformatics gene cloning prokaryotic expression

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Area of Science:

Molecular Biology
Plant Biochemistry
Medicinal Plant Research

Background:

GDP-mannose is crucial for synthesizing Codonopsis pilosula polysaccharides.
Phosphomannomutase (PMM) catalyzes a key step in GDP-mannose synthesis.

Purpose of the Study:

To clone and characterize the PMM gene from C. pilosula (CpPMM).
To investigate the correlation between CpPMM gene expression and polysaccharide synthesis in C. pilosula.

Main Methods:

Gene cloning using specific primers based on transcriptome data.
Bioinformatics analysis for gene and protein prediction.
Prokaryotic expression of the CpPMM gene.
Quantitative reverse transcription PCR (qRT-PCR) for gene expression analysis.

Main Results:

The full-length CpPMM gene (741 bp ORF, 246 amino acids) was cloned and found to be homologous to PMM from other species.
Bioinformatics predicted CpPMM as a hydrophilic, non-transmembrane protein located in the cytoplasm, belonging to the HAD superfamily.
Prokaryotic expression yielded a ~29 kDa fusion protein, primarily as inclusion bodies.
qRT-PCR revealed spatiotemporal expression patterns, with significantly higher CpPMM expression during the fruiting period, correlating with increased polysaccharide content.

Conclusions:

The CpPMM gene was successfully cloned and its prokaryotic expression system constructed.
CpPMM gene expression levels are highly correlated with C. pilosula polysaccharide content.
This study provides a foundation for further research into CpPMM function and polysaccharide biosynthesis pathways in medicinal plants.