Cloning and prokaryotic expression of transcription factor CtMYB1 gene in Carthamus tinctorius
10.7501/j.issn.0253-2670.2015.17.017
- Author:
Li-Li GUAN
1
Author Information
1. Bioreactor and Drug Development Reseach Center, Ministry of Education, Jilin Agricultural University
- Publication Type:Journal Article
- Keywords:
Carthamus tinctorius L.;
CtMYB1;
MYB;
Prokaryotic expression;
RACE;
RT-PCR;
Safflor yellow
- From:
Chinese Traditional and Herbal Drugs
2015;46(17):2603-2609
- CountryChina
- Language:Chinese
-
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.
