Isolation and bioinformatics study of TbJAMYC transcription factor involved in biosynthesis of Taxol from Iranian yew

Authors

1 Ph.D student of medical biotechnology, Medical Biotechnology Department, School of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran.

2 Corresponding author, Assist. Prof., Faculty of Agricultural and Natural Sciences, Biotechnology Department, Imam Khumaini International University, Qazvin, I.R. Iran. Email: nayeri@alumni.ut.ac.ir,

Abstract

  An anti-cancer drug called Taxol, or paclitaxel is one of the secondary metabolites of yew
tree (Taxus bacatta). Taxol is one of the most important and most useful anticancer drugs in the
treatment of lung cancer. Taxol prevents the depolymerization of microtubules and inhibits cell
division in tumor tissues. JAMYC is a transcription factor from MYC family with a DNAbinding

motif called bHLH. The gene involved in Taxol biosynthesis has been identified in
other yew species. The aim of this study was isolation and cloning of TbJAMYC gene and
bioinformatics analysis of its structure. Initially to obtain yew seedling, embryos were removed
from seeds of the species and cultured on MS medium containing activated charcoal and
ascorbic acid. Total RNA was extracted from the seedlings and cDNA was synthesized from
extracted RNA. The gene of TbJAMYC was amplified by PCR from cDNA. Amplified fragment
cloned in pTG 19 vector using TA cloning method and sequenced. Bioinformatics analysis was
carried out by MODELLER and Pfam softwares. The cloning was confirmed by endonuclease
digestion and sequencing. TbJAMYC bioinformatics analysis was carried out by Pfam and
MODELLER softwares that revealed three-dimensional structure of the DNA binding domain.
These analyzes showed that TbJAMYC protein is a member of MYC transcription factors
family that binds to DNA and regulates gene expression by bHLH domain. TbJAMYC
transcription factor involved in the biosynthesis of valuable anticancer Taxol drug. Hence Taxol
biosynthesis pathway manipulation via genetic engineering can be a new way to increase
production of the metabolite in yew plant.

Keywords

References

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