为阐明木本植物中核转录Y因子B亚基蛋白(NF-YB)及其编码基因在抗旱转录调控中的机制,运用生物信息学知识和方法,采用PCR克隆技术,以模式树种毛果杨基因组核转录Y因子B亚基蛋白同源序列为参考,从胡杨叶组织基因组材料中反转录获取了核酸长度为543 bp的目标基因PeNF-YB1.该基因编码的蛋白具有螺旋-环-螺旋二级结构,含有NF-YB蛋白保守结构域和功能氨基酸,不合核定位信号肽;GFP亚细胞定位表明该基因在核中表达;PEG6000溶液干旱胁迫下,该基因表达具有早期上调、延后转为下调的响应变化.研究认为克隆的PeNF-YB1是植物NF-YB亚基基因家族成员,并在干旱早期响应过程中起转录表达作用.研究结果不仅说明了NF-YB转录因子在木本植物中的抗旱作用,也为树木抗旱调控机制的阐明提供了重要参考.
The aim was to elucidate the transcriptional regulation mechanism of NY-YB proteins and their encoding genes from woody plants in drought resistance, the knowledge and tools of bioinformatics and PCR technology were applied for cloning a target sequence with 543 bp in length, named PeNF-YB1, from leave issues of Populus euphractica via reverse transcription approach, with the NF-YB homologue sequence in the genome of P. ttichocarpa as reference. The protein encoded by the gene possessed a helix-loop-helix secondary structure, conserved domains and functional amino acids of NF-YB protein. The protein did not have a nuclear localization signal peptide. The gene was observed to be expressed in unclear field with GFP subcellular localization method. Under drought stress on P. euphratica imposed by PEG 6000 solution, the gene was observed to be an up-regulated expression in early stage of treatment, and a lagging down-regulated expression. The studies suggested that the cloned PeNF-YB1 gene is a member of plant NF-YB gene and takes a regulative role in early drought response. The results of this study could not only verify the family role of NF-YB in wood plants resistance to drought, but also provide a basic clue for elucidating regulation mechanism of tree resistance to drought.