Cloning and prokaryotic expression of transcription factor CtMYB1 gene in Carthamus tinctorius / 中草药

ABSTRACT

Objective: To obtain a transcription factor gene MYB, clone a CtMYB1 gene from the safflower petals of Carthamus tinctorius, and performe its sequence analysis and prokaryotic expression vector construction. Methods: According to high expression Unigene123993 sequence in safflower transcriptome, MYB gene was cloned from safflower by RT-PCR and RACE methods. The full-length cDNA sequences CtMYB1 gene as templates, the open reading frame (ORF) of cDNA sequences was obtained by PCR. Prokaryotic expression vector pEASY-E1-CtMYB1 was constructed and the expression in E. coli BL21 (DE3) was transformed. Results: A MYB gene was successfully cloned from the safflower petals of C. tinctorius, named CtMYB1 (GenBank accession No. KJ524853). A full length cDNA of CtMYB1 was 893 bp and ORF was 750 bp, encoding a protein of 249 amino acid. The prokaryotic expression vector was obtained. SDS-PAGE results showed that the molecular weight was 30000, same with the relative molecular weight of predicted protein. Conclusion: CtMYB1 is cloned from safflower and the prokaryotic expression vector is constructed, which preliminarily proves that the gene is successfully expressed in E. coli.