向日葵肌动蛋白基因的cDNA克隆及进化分析  

Cloning and Evolutionary Analysis of Actin Gene from Sunflower(Helianthus annuus)

作　　者：李园莉[1] 江元清[1] 赵武玲[1] 阎隆飞[1] 

机构地区：[1]中国农业大学生物学院农业部植物生理生化重点开放实验室,北京100094

出　　处：《农业生物技术学报》2002年第1期67-71,共5页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金资助项目(No.39770052;39730280;39870077);所得序列在GenBank中的检索号为AF282624

摘　　要：本研究以向日葵（Helianthus annuus）为材料,提取其总RNA,根据植物肌动蛋白基因编码区的5＇和3＇末端设计简并引物。利用RT-PCR技术,用5＇RACE试剂盒扩增出向日葵肌动蛋白基因编码区全长。以豌豆肌动蛋白cDNA作探针的Southern杂交检测表明扩增出了肌动蛋白基因。将所获片段克隆到pGEM-T载体后进行测序,所得序列与GenBank已知肌动蛋白基因序列的相似性均在60％以上,其中与之相似性最高的锦葵为85％;与主要高等植物肌动蛋白氨基酸序列的相似性达80％以上。根据氨基酸序列的相似性绘制主要高等植物肌动蛋白的系统树（种系树）,表明向日葵肌动蛋白与锦葵肌动蛋白可能由同一祖先进化而来。Actin is highly conserved and plays an important role in many biological processes. Total RNA was isolated from sunflower(H. annuus)seedlings. Primers were designed according to actin coding region of plants, RT-PCR technique was employed by 5' RACE kit to obtain the actin coding region of sunflower.Southern blotting analysis by using probe from pea tendril actin gene coding region showed that there was highly similarity between actins of sunflower and pea tendril's.Putative actin gene was sequenced after cloned into pGEM-T vector and compared with other actin sequences in the GenBank.Our results showed that the similarity between the obtained sunflower actin nucleotide sequence and other known actin cDNA sequence was over 60%,and 85% with that of mallow. Also similarity of amino acid sequences between sunflower actin and those of others was over 80% . Based on the similarity of amino acids, phylogenetic tree of actins from higher plants was reconstructed, which shows that actin of H.annuus and actin of M. pusill a likely evolved from a common ancestor.

关 键 词：向日葵 肌动蛋白基因 快速扩增CDNA末端 系统树 克隆 进化分析 

分 类 号：Q946.1[生物学—植物学] Q943