Identification and recombinant expression of Crocin synthesis isoform gene in Saffron stigma

The existence of isoform genes in plants led to the creation of the different types of isomeric metabolites like these glycosylated forms. This study was conducted to investigate the identification and heterologous expression of Saffron stigmas Glycosyl transferase coding isoform gene during the pollination stage. The results have been obtained by sequencing and bioinformatics analysis of an isolated gene from saffron genomes by degenerate oligo’s revealed that the gene is in1283 bp length and belongs to CsUGT protein family which has Apo- plastic secretion in the cell. to evaluate the enzymatic function, firstly the isolated sequence was sub-cloned under arabinose induce promoter in pThio-UGT expression vector by Gibson assembly technique, then the recombinant vector transformed into BL21-pGro7 bacteria which were able to express chaperon proteins with EL& ES subunits. Followed by destructing the bacteria cell wall via ultrasound, the pellet was suspended by PBS solution and then the soluble proteins were extracted by boiling method. Finally, the protein electrophoresis by SDS pages10% was showed that the recombinant protein of CsUGT expressed correctly in bacteria with 69/5 kDa molecular weight. The gained results in this project could be applied to determine the breeding's strategies to improve qualitative and quantitative traits such as color and aroma in saffron.