Identification and Recombinant Expression of Crocin Synthesis Isoform Gene in Saffron Stigma

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HOSSEINPOUR AZAD NORADDIN

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Journal: Saffron Agronomy and Technology Year:2020 | Volume:7 | Issue:4 Page(s): 481-490

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Information Journal Paper

Title

Identification and Recombinant Expression of Crocin Synthesis Isoform Gene in Saffron Stigma

Author(s)

HOSSEINPOUR AZAD NORADDIN | Issue Writer Certificate

Keywords

Recombinant expressionQ1

Glycosyl transferaseQ1

CrocinQ1

E.coliQ2

Abstract

The existence of isoform genes in plants led to 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 the 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 the 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 the boiling method. Finally, the protein electrophoresis by SDS pages 10% showed that the recombinant protein of CsUGT expressed correctly in bacteria with 69. 5 kDa molecular weight. The results obtained 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.

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APA: Copy

HOSSEINPOUR AZAD, NORADDIN. (2020). Identification and Recombinant Expression of Crocin Synthesis Isoform Gene in Saffron Stigma. SAFFRON AGRONOMY AND TECHNOLOGY, 7(4 ), 481-490. SID. https://sid.ir/paper/375923/en

Vancouver: Copy

HOSSEINPOUR AZAD NORADDIN. Identification and Recombinant Expression of Crocin Synthesis Isoform Gene in Saffron Stigma. SAFFRON AGRONOMY AND TECHNOLOGY[Internet]. 2020;7(4 ):481-490. Available from: https://sid.ir/paper/375923/en

IEEE: Copy

NORADDIN HOSSEINPOUR AZAD, “Identification and Recombinant Expression of Crocin Synthesis Isoform Gene in Saffron Stigma,” SAFFRON AGRONOMY AND TECHNOLOGY, vol. 7, no. 4 , pp. 481–490, 2020, [Online]. Available: https://sid.ir/paper/375923/en

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