Cloning and Prokaryotic Expression of Arabidopsis AtTCP4 Gene

doi:10.7668/hbnxb.20192154

Abstract

Abstract: To investigate the function of AtTCP4 downstream target genes and in plant growth and development. Bioinformatics analyzed transcription factor TCP4 family and its protein sequence.The prokaryotic expression vector was constructed and the protein was induced and purified. EMSA was done to analyze AtTCP4 protein can bind to the cis-acting elements in the promoter region of LOX2 gene.The bioinformatics analysis showed that the transcription factor TCPs of Arabidopsis thaliana contain a conserved bHLH domain, and the phylogenetic tree showed that AtTCP4 belonged to the class Ⅱ TCP protein. The prediction of protein tertiary structure indicated that TCP4 could form homodimer or heterodimer. Specific primers were used to obtain the CDS sequence of AtTCP4 gene from Arabidopsis thaliana, whose open reading frame (ORF) was 1 263 bp. The prokaryotic expression vector pET-28a-AtTCP4 was constructed and transformed into Rosetta (DE3) competence. After IPTG induction, protein purification and Western Blot detection proved that the AtTCP4 protein was obtained. EMSA experiment verified that the purified AtTCP4 protein could bind to the cis-acting elements of gene LOX2. It affected biosynthesis of plant hormone jasmonic acid and regulated the process of plant growth and development.

Key words: Arabidopsis thaliana, Transcription factor, TCP, Gene cloning, Prokaryotic expression, Protein purification

CLC Number:

LEI Qidong, SUN Xudong, XU Huini. Cloning and Prokaryotic Expression of Arabidopsis AtTCP4 Gene[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(5): 24-28. doi: 10.7668/hbnxb.20192154.

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