CAJ | 학술논문

通过筛选百脉根(Lotus 7aponicus)基因组文库,克隆了LjCYC1(Lotus japonicus Cycloidea-like1)基因.L7CYC1是金鱼草(Antirrhium)CYC (Cycloidea)基因的同源基因,CYC属于TCP[TBl(teosinte branchedl),CYC,PCFs(PCFl and PCF2)]基因家族,编码转录调控子控制金鱼草花的对称性.基因组序列分析表明,LjCYC1的开放阅读框(ORF)由两个外显子和一个内含子组成,其cDNA编码的LjCYC1蛋白包含了370个氨基酸.蛋白序列分析显示,LjCYC1包含一个TCP结构域和一个R结构域,属于TCP结构域蛋白家族的CYC/TBl亚家族;LjCYCl氨基酸序列与CYC相比,一致性和相似性分别为39.0%和42.6%.不同长度的LjCYCl-cDNA与报告基因GUS融合后,通过粒子轰击(particle bombardment)方法在洋葱表皮细胞瞬时表达融合蛋白,观察到包含了TCP结构域的融合蛋白能够进行细胞核定位,提示LjCYCl可能作为转录因子行使功能;TCP结构域自身不能完成核定位过程,还需要结构域两侧旁邻氨基酸序列的协助.
통과사선백맥근(Lotus 7aponicus)기인조문고,극륭료LjCYC1(Lotus japonicus Cycloidea-like1)기인.L7CYC1시금어초(Antirrhium)CYC (Cycloidea)기인적동원기인,CYC속우TCP[TBl(teosinte branchedl),CYC,PCFs(PCFl and PCF2)]기인가족,편마전록조공자공제금어초화적대칭성.기인조서렬분석표명,LjCYC1적개방열독광(ORF)유량개외현자화일개내함자조성,기cDNA편마적LjCYC1단백포함료370개안기산.단백서렬분석현시,LjCYC1포함일개TCP결구역화일개R결구역,속우TCP결구역단백가족적CYC/TBl아가족;LjCYCl안기산서렬여CYC상비,일치성화상사성분별위39.0%화42.6%.불동장도적LjCYCl-cDNA여보고기인GUS융합후,통과입자굉격(particle bombardment)방법재양총표피세포순시표체융합단백,관찰도포함료TCP결구역적융합단백능구진행세포핵정위,제시LjCYCl가능작위전록인자행사공능;TCP결구역자신불능완성핵정위과정,환수요결구역량측방린안기산서렬적협조.
The LjCYC1 (Lotus japonicus Cycloidealike l) gene, a homolog of CYC(Cycloidea) belonging to the TCP [TBl(teosinte branched l),CYC, PCFs (PCFl and PCF2)] gene family and encoding a predicted transcription factor and being proposed controlling different aspects of plant development, was isolated from the papilionaceous plant Lotus japonicus by screening the genomic DNA library, in order to test the functional conservation and divarication of CYC-like genes in legume. Sequence analyses indicate that LjCYC1 gene contains two exons and one intron and encodes a 370-AA peptide LjCYC1. The putative protein, LjCYC1, contains a TCP domain and an R domain, being a member of the CYC/TB 1 subfamily of TCP family, and has 39.0% identity with and 42.6% similarity to CYC. LjCYC1-cDNA was cloned through RT-PCR. Different regions of the LjCYClcDNA were fused with the report gene GUS and then the fused constructs were transiently expressed in the onion epidermal cells through particle bombardment. Results of GUS and DAPI staining showed that the chimeric proteins with TCP domain were localized within the nucleus, confirming that LjCYC1 may act as a transcription factor. But the TCP domain itself could not confer the nuclear localization because the chimeric proteins with TCP domain alone were dispersed all over the transformed cells.