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根据彩叶草叶片小型EST库中一条具有1个富亮氨酸重复(leucine-rich repeat,LRR)结构域的EST序列,采用RACE与文库结合的方法,克隆了1个具有5个LRR结构域的全长cDNA,SsLRP(LRR-Related Protein)(GenBank登录号FJ787729)。SsLRP cDNA全长1024bp,包含一个657bp的ORF框,编码218个氨基酸。其5’-UTR区含有2个终止子TAG,3’-UTR区具有推测的加尾信号AATAAA。SsLRP蛋白N端具有的信号肽和保守的亮氨酸拉链结构域,具有5个保守的LRR结构域,多个磷酸化位点和N-糖基化位点。多序列比对和系统进化分析表明,SsLRP与番茄SlLRP同源性最高。二级结构和三级结构预测表明,SsLRP的功能可能与保守的LRR结构域密切相关,推测该基因可能参与蛋白间的相互作用与信号识别。RT-PCR分析表明,SsLRP与番茄SlLRP具有相似的表达模式,在正常植株的根、茎、叶和花中都有表达,在受菌核病感染植株的茎和叶中表达上调。
Based on a EST sequence with a leucine-rich repeat (LRR) domain in a small EST library of C. folium leaves, a library of 5 LRR domains Full length cDNA, SsLRP (LRR-Related Protein) (GenBank Accession No. FJ787729). SsLRP cDNA is 1024 bp in length and contains a 657 bp ORF encoding 218 amino acids. Its 5’-UTR region contains two terminator TAGs, and the 3’-UTR region has a putative tailed signal, AATAAA. The signal peptide possessed by N-terminal of SsLRP protein and the conserved leucine zipper domain have 5 conserved LRR domains, multiple phosphorylation sites and N-glycosylation sites. Multiple sequence alignment and phylogenetic analysis showed that SsLRP shared the highest homology with tomato SlLRP. Secondary structure and tertiary structure prediction show that the function of SsLRP may be closely related to the conserved LRR domain, suggesting that the gene may be involved in protein interactions and signal recognition. RT-PCR analysis showed that SsLRP has a similar expression pattern with tomato SlLRP, which is expressed in roots, stems, leaves and flowers of normal plants, and is up-regulated in stems and leaves infected with sclerotinia.