CAJ | 학술논문

Dof（DNA binding with one finger）转录因子是一类植物特有的、进化上高度保守的基因家族，对植物多方面的生理过程均发挥调控作用。根据Dof家族中的一个B06基因的cDNA保守序列设计引物，对8个品种花生的B06基因进行PCR扩增克隆相似基因，并测定序列。实验成功克隆出四粒红和073103的完整1306基因和其余6个品种的大部分B06基因，其中073103含有一个内含子，其余7个品种不合内含子。通过比较8个品种间B06基因的序列差异与品种性状间的关系，并通过生物信息学对基因及编码蛋白进行分析，探究B06蛋白可能的功能。结果发现：典型的B06蛋白由176个氨基酸组成，在N-端附近37个氨基酸区域处形成保守的单锌指DOF结构；推测在DOF区域后的59—67氨基酸处有核定位信号序列；蛋白序列中有多个N-糖基化、蛋白激酶C磷酸化、酪蛋白激酶Ⅱ磷酸化、N-十四烷酰基化位点，可以接受不同途径的信号刺激，修饰前后发生定位或活性的变，推测1306蛋白可能与植物对外界刺激的反应相关。结合性状分析，推测1306蛋白与花生种子中的脂肪和蛋白质含量高低无明显联系。试验为进一步深入研究B06蛋白的功能奠定了基础。
Dof（DNA binding with one finger）전록인자시일류식물특유적、진화상고도보수적기인가족，대식물다방면적생리과정균발휘조공작용。근거Dof가족중적일개B06기인적cDNA보수서렬설계인물，대8개품충화생적B06기인진행PCR확증극륭상사기인，병측정서렬。실험성공극륭출사립홍화073103적완정1306기인화기여6개품충적대부분B06기인，기중073103함유일개내함자，기여7개품충불합내함자。통과비교8개품충간B06기인적서렬차이여품충성상간적관계，병통과생물신식학대기인급편마단백진행분석，탐구B06단백가능적공능。결과발현：전형적B06단백유176개안기산조성，재N-단부근37개안기산구역처형성보수적단자지DOF결구；추측재DOF구역후적59—67안기산처유핵정위신호서렬；단백서렬중유다개N-당기화、단백격매C린산화、락단백격매Ⅱ린산화、N-십사완선기화위점，가이접수불동도경적신호자격，수식전후발생정위혹활성적변，추측1306단백가능여식물대외계자격적반응상관。결합성상분석，추측1306단백여화생충자중적지방화단백질함량고저무명현련계。시험위진일보심입연구B06단백적공능전정료기출。
Dof（DNA binding with one finger）transcription factors is a highly conserved gene family, which play a role of regulation and control in various physiological processes of plant. According to conserved sequences of 1306 gene eDNA, with a pair of primers, DOF 1306 similar genes were cloned, sequenced and compared from genomic DNA of eight cultivar. The results showed that a typical B06 protein is composed of 176 animo adds, in which a DOF domain close to N-terminal with 37 animo acids forming the typical single zinc finger structure; B06 protein does not contain a signal peptide at the N-terminal, and the domain next to DOF domain with 9 animo acids are predicted as a nuclear localization signal （NLS）; there exit 3 N-glycosylation sites, 2 protein kinase C phosphorylation site, 1 casein kinase II phosphorylation site and 3 N-myristoylation sites acceptting signal stimulus via different ways, suggest that localization and activity of B06 alters before or after the modification; presumably related with the responds to the outside stimulus.