Molecular cloning and expression analysis of cinnamic acid 4-hydroxylase gene from Isatis indigotica / 中草药

ABSTRACT

Objective: To clone cinnamate 4-hydroxylase (C4H) gene from Isatis indigotica and to analyze its bioinformatics and expression. Methods: The full-length cDNA of C4H was 1 674 bp (GenBank accession No. GU014562) long with an open reading frame (ORF) of 1 530 bp encoding a polypeptide of 509 amino acid residues. The comparison of C4H genomic DNA sequences and C4H cDNA sequence revealed that the C4H genomic DNA contained two introns. Southern-blotting analysis indicated that C4H was a multiple copy gene. C4H expression could be detected in all tissues at different expression levels, with the strongest expression in the roots. Further expression analysis revealed that the signaling components of defense/stress pathways, such as ultraviolet-B radiation (UV-B), methyl jasmonate (MeJA), abscisic acid (ABA), and Gibberellins (GA3) could up-regulate the C4H transcript levels compared with the control. Conclusion: We have first extracted the full length cDNA of C4H gene from I. indigotica, and its structural and bioinformatic analyses are carried out, which will help us to further illuminate this pathway. The research also provides a possibility to study the antiviral active constiuents in I. indigotica by plant secondary metabolic engineering.