Cloning and Expression Vector Construction of One CBF-like Gene from Poncirus

摘要

CRT/DRE binding factor(CBF )/DREB1 are one of the most important transcript factors that regulate expression of low temperature related genes in plant. So cloning and analysis CBF gene, which is the key gene of plant cold-resistance, is of great significance. To in-depth study the function of the CBF gene, we designed one pair of specific primers, and obtained a full-length CBF gene using RT-PCR, named Ptcbfb. An overexpression vector containing the cloned CBF gene was constructed by place the CaMV55s promoter and the downstream Nos terminator. RNAi interfering expression vector was also constructed using part of the full-length CBF gene fragment and the interfering vector DHRVP, and the part fragment of CBF gene and its inverted fragment were placed in the vector head-to-head with an intron sequence of the vector located in the middle of them. The two vectors were transferred into Agrobacterium tumefaciens EHA105. This is the foundation of citrus genetic transformation and further study.