Cloning of flavanone 3-hydroxylase gene from Lithocarpus polystachyus and its sequence analysis / 中草药

ABSTRACT

Objective: To clone flavanone 3-hydroxylase (F3H) gene from Lithocarpus polystachyus, and to understand its gene characteristics and initially investigate its expression level in different organs. Methods: The total RNA and genomic DNA from blade of L. polystachyus were extracted. Based on the result of RNA-seq, a pair of specific primers were designed. cDNA and DNA sequences of F3H gene from L. polystachyus were amplified by PCR, then bioinformation analysis was performed after sequencing. The expression level of F3H gene in different organs of L. polystachyus was detected by qRT-PCR. Results: The full length of cDNA of F3H gene was 1 340 bp containing a 1 092 bp open reading frame that encoded 393 amino acids, and F3H was located in the cytoplasm. The result of qRT-PCR showed that F3H gene expressed in different organs of L. polystachyus, and the expression levels of F3H gene were significantly different in different organs (P < 0.05). Conclusion: The F3H gene of L. polystachyus was cloned and its bioinformation was analyzed for the first time, proving that the expression level of F3H gene in different organs of L. polystachyus was different. This finding lays a foundation for the studies on secondary metabolism of flavonoids in L. polystachyus.