Cloning, bioinformatics, expression mode analysis of crocetin glycosyltransferase UGTCs4 / 中草药

ABSTRACT

Objective: With the aim to obtain crocetin glycosyltransferase UGTCs4 in cultured saffron suspension cell, and carry out its bioinformatics and expression mode analysis. Methods: A homologous cloning strategy and 5’ RACE methods were adopted, the full-length cDNA sequence of a crocetin glycosyltransferase, designated UGTCs4 (GeneBank number KX398932), was obtained. The characteristics of physiochemical properties, structure and function of the deduced UGTCs4 protein were determined using a series of bioinformatics tools. Semi-quantitative PCR was used for gene expression analysis. Results: The results showed that the full length cDNA of UGTCs4 was 1 380 bp in length and encoding a 459 amino acid; UGTCs4 had high identities (83.2%) with UGTCs2 protein from saffron; UTGTCs4 had the same evolutionary tree as UGTCs2. UGTCs4 transcripts were constitutively expressed in the leaves, stems, and roots. UGTCs4 gene could respond to multiple treatments of indoleacetic acid (IAA), abscisic acid (ABA), gibberellin (GA), hydrogen peroxide (H2O2), and methyl jasmonate (MJA), which promoted its transcription. Conclusion: cDNA of crocetin glycosyltransferase was cloned from suspension cells for the first time and the response of UGTCs4 to different inducers was confirmed. Molecular characterization of UGTCs4 will be useful for further functional determination of the gene involving in the crocin biosynthesis and expression regulation.