摘要
通过筛选百脉根(Lotus japonicus)基因组文库,克隆了UjCYC1(Lotus japonicus Cycloidea-like1)基因。LjCYC1是金鱼草(Antirrhium)CYC(Cycloidea)基因的同源基因,CYC属于TCP[TB1(teosinte branched 1),CYC,PCFs(PCF1 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/TB1亚家族;LjCYC1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LjCYC1-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LjCYC1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。
The LjCYC1 (Lotus japonicus Cycloidealike 1) gene, a homolog of CYC(Cycloidea) belonging to the TCP [TB1(teosinte branched 1),CYC, PCFs (PCF1 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 LjCYC1-cDNA 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 LjCYC 1 may actas 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.
出处
《植物生理与分子生物学学报》
CAS
CSCD
2004年第5期523-532,共10页
Journal Of Plant Physiology and Molecular Biology
基金
国家自然科学基金(No.39825102)~~
关键词
百脉根
LjCYCl
TCP基因家族
融合蛋白
粒子轰击
Lotus japonicus
LjCYCl (Lotus iaponicus Cycloidea-like 1)
TCP gene family
fused protein
particle bombardment
