莲藕CBF2基因cDNA克隆及序列分析

Cloning and Sequence Analysis of CBF2 in Lotus Loot( Nelumbo Nucifera Gaertn. )

目的:克隆莲藕CBF2基因的c DNA全长并对其进行序列分析。方法:根据已有的ESTs序列,设计3'和5'端RACE引物,运用RACE技术克隆莲藕CBF2基因c DNA全长。结果:克隆得到全长为1 560bp c DNA序列,其中包括350bp的3'非编码区和124bp的5'非编码区及一个1 086 bp的完整开放阅读框,编码362个氨基酸,序列末端有poly(A)尾。其核苷酸序列与NCBI数据库中大豆DREB2C/CBF2、马铃薯DREB2A/CBF2、黄瓜DREB2A/CBF2及花生DREB2A/CBF2的同源性较高,分别为83%、77%、76%和76%,因此把该基因命名为Lr CBF2。氨基酸序列进化树分析表明该基因与葡萄相关基因亲缘关系较近。结论:分离克隆得到莲藕CBF2基因,为进一步研究莲藕中该基因的功能奠定基础。

Objective： Cloning and sequence analysis of full- length CBF2 gene in lotus root. Method： Full- length CBF2 gene was isolated from lotus root according existed ESTs sequence using 3＇and 5＇RACE technique. Result： Full- length CBF2 gene was 1 560 bp with poly（ A）,containing 350 bp 3＇non- coding region,124 bp 5＇non- coding region and a 1 086 bp open reading frame which encoded putative 362 amino acids. Compared against NCBI database,this gene showed 83%,77%,76% and 76% homolog with DREB2 C / CBF2 from soybeanm,potato and cucumber respectively. Therefore,this gene was designated as Lr CBF2. In addition,the result of hylogenetic analysis indicated that Lr CBF2 showed a closer genetic relationship with that of grape. Conclusion： A CBF2 gene was cloned from lotus root in his experiment with aim to exploring gene function in future.