烟草FIE和MSI1基因的克隆及序列分析

采用电子克隆与实验克隆结合的方法获得了烟草胚乳发育相关基因NTFIE和NTMSI1的cDNA序列,序列号分别为EU375458和EU375459.序列分析结果表明,这两个cDNA序列均含有完整的开放读码框,分别编码370和424个氨基酸,含有保守的WD基序.氨基酸序列比对和系统发育分析结果显示,不同物种之间FIE和MSI1基因编码氨基酸序列同源性都较高.组织表达分析结果表明,这两个基因均具有一定程度的组织表达特异性,NTFIE cDNA基因在花中的表达量最多,但在根和茎中未检测到表达,而NTMSI1 cDNA基因只在离体培养的细胞和根中特异性表达.

Cloning and Sequence Analysis of FIE and MSI1 Genes from Nicotiana tabacum

cDNA sequences of endosperm development related genes,NTFIE and NTMSI1,were cloned by using in silico cloning combined with experimental RT-PCR from Nicotiana tabacum.Sequence analysis showed that both of the two cDNAs have intact open reading frame (ORF),encoding 370 and 424 amino acids,respectively.They both contain WD-motif.Protein multiple-alignment and phylogenetic analysis suggested that amino acids sequences encoded by the NTFIE and NTMSI1 show strong similarity in different species,indicating that these two genes were conserved in plants.The times of differentiation of NTFIE and NTMSI1 are in accord with their classical categories.Tissue specific expression analysis with UniGene database showed that,to some extent,these two genes have tissue specific expression.NTFIE has the highest expression in flowers but not expressed in root or stem.NTMSI1,on the other hand,only expressed in cell culture and root.