Cloning And Expression In Prokaryocyte Of Transcription Factor CBF1 And NAC Gene In Arabidopsis Thaliana

Transcription factors (transcription factor, TF) is a type of protein that has special structure and functioning in regulation of gene expression, also known as trans factors. By identifying the cis acting elements in the promoter of eukaryotic genes, the expression of the target gene is regulated precisely. Arabidopsis thaliana is a model plant for genetics research. It is a good material for the study of function, evolution and ecology in the field of plant biology and genetics. The advantage of the Arabidopsis for plant functional gene study is easy to obtain. Transcription factors can regulate the expression of target genes in the process of plant functional gene expression, in order to express target genes in a certain time and space with certain levels. The expression of many genes in plant is controlled by specific transcription factors and specific cis acting elements. CBF1 and NAC are two important activate factors for the expression of stress-resistant genes in plants. CBF1 transcription factor can regulate the expression of a group of anti-drought and low temperature genes in Arabidopsis, which can effectively improve the resistance of plants to drought and low temperature, and NAC is mainly involved in the regulation of plant resistance to low temperature, drought, high salt and other related factors.In this study, we studied the function of NAC and CBF1 by gene cloning and expression using thaliana(Arabidopsis) leaves as the research material. Trizol reagent was used to extract total RNA from leaves, and reverse transcribed to cDNA, then the CBF1 gene and NAC gene were amplified. Nextly the target genes were cloned and identified by enzyme digestion and sequencing. Finally, the sequence was analyzed and the corresponding protein was translated into DNAMAN. The water affinity of proteins were predicted. Finally CBF1 gene was used to construct the prokaryotic expression vector, and the expressed protein was identified by differentially expression profile. SDS-PAGE results showed that the expressed protein of CBF1 gene was 24 kD. The preliminary results of the study provide a reference for the study of the plant’s stress-resistance, also support the next step study that use the above two genes to obtain new plant strains with strong resistance.