Cloning Of CBF1, CBF4 Genes And Promoter In Arabidopsis Thaliana And Construction Of Plant Expression Vectors

The key genes CBF1,CBF4 on this experiment study are the important members of the small CBF gene family. The CBF gene family is a small gene family which includes CBF1,CBF2,CBF3,CBF4 genes. They play a very important part in plants'resistance salt and alkali mechanisms in plants.Plants'CBF transcription activators play an exceedingly important part in plants'growth and development and the response to external environment. The CBF transcription activators are one of the Trans-acting Factors which Specific induced by low temperature. They can combine with the regulatory element CRT/DREDNA(C-repeat/dehydration-responsive element(DRE)DNA binding protein), Promote the low temperature and dehydration induced genes'whose promoter including the regulatory element expression,thus activated the various anti-adversity mechanisms in plants.To improve the stress resistance of in alfalfa, genes of CBF1,CBF4 transcription factors and its promoter were isolated from Arabidopsis thaliana by PCR method.The plant expression vectors of PBI121-CBF1,PBI121-CBF4,PBI121-CBF1-PRO and PBI121-PRO-CBF1 were constructed and confirmed by double enzyme digestion.The research got the preliminary results as follows:1. The research selected the primerâ… which can isolate the gene of CBF1 by PCR method; Selected the primerâ…¢which can isolate the promoter of CBF1 by PCR method; Selected the primerâ…¤which can isolate the gene of CBF4 by PCR method. 2. genes of CBF1,CBF4 transcription factors and its promoter were isolated from Arabidopsis thaliana by PCR method. 3. The plant expression vectors of PBI121-CBF1,PBI121-CBF4,PBI121-CBF1-PRO and PBI121-PRO-CBF1 were constructed and confirmed by double enzyme digestion.