Isolation, Cloning, and Bioinformatic Analysis of UGT Gene in Saffron (Crocus sativus L.) of Iran

Saffron is the most expensive spice in the world. The economic value of saffron is due to the existence of apocarotenoids in its stigma. Therefore, the isolation and characterization of genes involving in apocarotenoids metabolism is on particular importance. In this research, beacause the importance of CsUGT gene in crocin biosynthesis, it was isolated, cloned the E.coli strain DH5α. The full length gene was sequenced and registered in the NCBI. In order to characterize CsUGT gene, first the protein sequence was obtained, and then the physical and chemical characteristics and physiology of the CsUGT protein were analyzed by Protparam, SOPMA, ProtScale, Pfam, ProtComp, SignalP, TMHMM and ChloroP servers and tools. Also, using the Swiss-Model server, the 3D structure of this protein was investigated and Ramachandran diagram was drawn to validate the 3D drawn model structure. According to the results, CsUGT protein with 462 amino acids has the conserved sequence of glycosyltransferase family proteins and was identified as a polar protein, stable at high temperatures and without hydrophobic domain. CsUGT protein has no peptide signal or binding signals and has a cytoplasmic location. This research made it possible to isolate the CsGTS gene of saffron and optimized the conditions of its transfer into a vector. In addition, the results of CsUGT protein structure analysis provide the basis for future functional studies and can also provide valuable information regarding the behavior and reaction of this enzyme in the synthesis of saffron apocarotenoids. In addition, these results can be useful in the future programs of Iranian saffron genetic modification.