摘要
CBF是植物冷驯化过程中的关键性调节因子。利用PCR及染色体步移法从新疆特有拟南芥近源种植物———小拟南芥的基因组中克隆了CBF基因的同源序列(DQ207404)。生物信息分析表明,该序列具有完整的读码框,推定的蛋白质序列与拟南芥CBF1,2,3的相似性分别为:87.6%、86.6%和88.1%。以植物表达载体pBI121为基础,构建了由组成型启动子35S调控ApCBF的植物表达载体pCB111。通过根癌农杆菌介导,采用叶盘法转化烟草,PCR检测初步证明ApCBF基因已整合到烟草基因组中。为利用ApCBF基因改良植物抗逆性奠定了物质基础。
Transcription activator CBF is an critical regulator in cold acclimation of plant. An ortholog of CBF： ApCBF （DQ207404） was amplified and cloned by PCR and chromosome walking from the genomic DNA of the Arabidopsis pumila. It is special in Xinjiang. Sequence measurement and analysis showed that it included a complete open reading frame（ORF） ,the deuced protein sequences were 87.6% ,86.6% ,88.1% identical to the three Arabidopsis CBF1,2,3 respectively. The plants express vector pBI121 was digested and fused ApCBF with CaMV35S promoter to construct plant express vector pCB111. ApCBF gene was introduced into tobacoo by co-culturing excised leafs with Agrobacterium tumsfaciens. The trans-genic nature of the regenerants was demonstrated by plant growth on medium containing kanamycin. PCR analysis confirmed that ApCBF gene had been incorporated into the genome of tobacoo.
出处
《石河子大学学报（自然科学版）》
CAS
2006年第5期591-595,共5页
Journal of Shihezi University(Natural Science)
关键词
小拟南芥
CBF
基因克隆
烟草转化
Arabidopsis pumila
transcription activator CBF
gene cloning
tobacoo transformation
