Gene cloning of farnesyl pyrophosphate synthase in Atractylodes lancea and its expression pattern analysis / 中草药

ABSTRACT

Objective: To clone the full-length cDNA encoding farnesyl pyrophosphate synthase (FPPS) from Atractylodes lancea and analyze its expression. Methods: The full-length cDNA of FPPS in A. lancea was cloned via homology-based cloning and rapid amplification of cDNA ends approach. Also, the characterization of gene was revealed by bioinformatic analysis. The expression of FPPS was determined by qRT-PCR while the content of sesquiterpenes of rhizome in different growth stages in A. lancea was measured by GC-MS. Meanwhile the correlation between them was analyzed. Results: The full-length cDNA (1320 bp) of FPPS gene was obtained (AlFPPS, GenBank accession number KX443242), with an open reading frame of 1029 bp, encoding 342 amino acids. The deduced AlFPPS protein sequence contained five conserved motifs, two of which is full of Asp (DDXXD). qRT-PCR and GC-MS results showed a significant positive correlation between the content of sesquiterpenes and AlFPPS expression level. Conclusion: It can be primarily deduced that AlFPPS gene should be an important control point in the biosynthetic pathway of sesquiterpenes in A. lancea. This work provides scientific basis for clarifying the biosynthetic pathway of sesquiterpenes and application of biological engineering.