ग्लाइकॉमिक्स और लिपिडोमिक्स जर्नल
खुला एक्सेस

अमूर्त

Cloning, recombinant expression in Escherichia coli and DNA binding properties of Datura metel L. Myb transcription factor

Priya Arora

MYB proteins are a superfamily of transcription factors that play significant roles in developmental and defense-related processes in plants. The MYB family is characterized by the presence of a conserved DNA-binding domain called the MYB domain, which normally contains two or three imperfect repeat sequences. In the present study, we report the cloning, expression and DNA binding properties of Myb transcription factor from Datura metel L. An open reading frame of 856 bp was detected which could encode a protein of 278 amino acid residues. The cDNA encoding DmMyb was cloned in pRSETA expression vector and heterologously expressed in E.coli. The DmMyb expressed as histidine-tagged fusion protein was purified using Ni-NTA affinity chromatography. In addition, the ability of partially purified recombinant DmMYB protein to bind MBCE (Myb binding cis-elements) was evaluated with five known Myb specific cis-acting elements (MSCE 1, MSCE 2, MSCE 3, MSCE 4 and MSCE 5) using the electrophoretic mobility shift assay (EMSA). The binding assay indicated that the partially purified recombinant DmMyb interacts with three Myb-specific cis-acting elements (MBCE 3, MBCE 4 and MBCE 5). Further studies are required for providing deep insights into transcriptional regulation of Datura metel genes/gene networks which is associated with the DmMyb transcription factor