关键词: 葡萄, 葡萄, 热激转录因子, SSR, 分子标记

Abstract: The aims are to further study the function of CBF2 target protein, and investigate the molecular mechanism of cold resistance of CBF2 transcription factor in almond. The leaves of Xinjiang cultivated almond ‘Shuangruan’were used as materials. The CBF2 gene was cloned from the genomic DNA with PCR technique, and named AcCBF2. The recombinant prokaryotic expression vector pET- AcCBF2 was constructed byinserting the DNA fragment into the prokaryotic expression vector pET-32a(+), and then transformed into E. coli Rosetta(DE3). The bioinformatics results showed that the open reading frame was 717 bp and encoded a protein of 238 amino acids, its molecular weight was 26.59 kD and isoeletctric point was 5.29. Phylogenetic tree analysis showed that AcCBF2 had closest genetic relationship with the genes of almond CBF1, Prunus mira Koehne and Peach. The SDS-PAGE displayed that the weight of expressed proteins was 47 kD, which was consistent with the expected protein size. The optional induction condition of AcCBF2 gene in E. coli Rosetta was 4 h of induction in 0.2 mmol/L IPTG.