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CBF1转录因子能调控一组抗干旱、抗低温基因的表达,更有效地提高植物抗干旱、抗低温的能力。以抗寒性较强的长白山野生山丁子(耐-42℃低温)为材料,通过RT-PCR扩增出山丁子CBF1基因,随后亚克隆到pMD18-T载体,序列分析表明与基因数据库中山丁子CBF1对应区域相似性为100%,将该片段亚克隆到原核表达载体pBI121中构建原核表达质粒,转化大肠杆菌E.coli DH5α,在AMP抗性的LB平板上挑取重组质粒,进行PCR并测序结果与预期一致。
CBF1 transcription factor can regulate a group of anti-drought and anti-low-temperature gene expression, more effectively improve the plant’s ability to resist drought and low temperature. The CBF1 gene was amplified by reverse transcription polymerase chain reaction (RT-PCR) from the wild hardy C. chinensis (tolerated at -42 ℃), and then subcloned into pMD18-T vector. The sequence analysis showed that the CBF1 gene was co- The corresponding region was 100%. The fragment was subcloned into the prokaryotic expression vector pBI121 to construct a prokaryotic expression plasmid, which was then transformed into E. coli DH5α. The recombinant plasmid was picked on an LB-resistant AMP plate for PCR and sequencing As expected.