烟草ovate同源基因的全长cDNA克隆与序列分析

Cloning and Sequence Analysis of ovate Orthologous Gene in Tobacco (Nicotiana tabacum L.)

根据番茄中控制果实形状的主效数量性状基因ovate的序列,用生物信息学方法从茄科植物烟草中获得直系同源ovate基因(NTovate)的特异片段,经鉴定,此基因在烟草中至少有2个拷贝。在此基础上用cDNA末端快速扩增(RACE)方法,获得其中1个拷贝的1059bpNTovate全长cDNA序列。序列分析表明,NTovate cDNA序列编码352个氨基酸,其蛋白序列与番茄ovate蛋白序列和拟南芥ovate蛋白家族AtOFP7蛋白分别为70%和36%的序列一致率,而与此家族中其他蛋白以及水稻ovate蛋白仅在保守的ovate结构域有较低的同源性。此基因已在GenBank中登录(EU043369)。

Tomato ovate gene has been cloned and proved to control the fruit shape development. In this study, ovate orthologous gene in tobacco （Nicotiana tabacum）, which belongs to Solanaceae and doesn＇t have fruit development, was cloned and compared with homologs in tomato and other model species. Two sequence fragments with 90% identity were identified using bioinformatics methods. Two copies of tobacco ovate ortholog （NTovate） were then ,confirmed by sequence specific PCR. A 1 059 bp full length cDNA of NTovatel was obtained using rapid amplification of cDNA ends （RACE）. Further analysis on NTovatel cDNA indicated that it encoded a peptide containing 352 amino acids. The amino acid sequence comparison with tomato ovate protein and AtOFP7 （Arabidopsis thaliana ovate family protein 7） showed that the sequence identity was 70% and 36%, respectively. Other AtOFPs and ovate family protein-like in rice only showed homolog in the ovate domain with much lower sequence identity. The full length cDNA sequence of NTovatel was registered in GenBank with accession number EU043369.