Molecular cloning, differential expression, and bioinformatics analysis of G10H gene in Gentiana rigescens / 中草药

ABSTRACT

Objective: To clone the geraniol-10-hydroxylase (G10H) gene from Gentiana rigescens and analyze the gene expression. Methods: The cDNA sequence of G10H was obtained from G. rigescens using PCR technique. The physical and chemical properties, secondary structure and tertiary structure of G10H protein were forecasted and analyzed using related software. The expression of G10H gene was detected using real-time PCR in roots, stems, and leaves of G. rigescens. Results: The cloned G10H gene was 1 400 bp including 1 248 bp open reading frame and encoding a predicted protein of 415 amino acids. Bioinformatics predicted that the gene encoding protein molecular formula was C2131H3390N586O615S17, the isoelectric point was 7.62, the instability coefficient was 44.20, and the hydrophobic coefficient was -0.245. RT-PCR showed that G10H gene expressed in all organs. And the highest expression level was found in roots, while the lowest in the stems. Conclusion: This G10H gene is cloned from G. rigescens for the first time. The results provide a foundation for exploring the mechanism of gentiopicroside biosynthesis in G. rigescens.