甜菜Actin基因片段的克隆及序列分析

根据其它植物Actin基因设计一对简并性引物,以甜菜根总RNA为模板,采用RT-PCR的方法扩增出Actin基因片段并克隆到pUCm-T载体上,阳性克隆经PCR鉴定测序。序列分析表明,克隆得到Actin基因家族的两个成员,其片段长度均为598 bp,编码198个氨基酸,它们间的同源性为87%。将其分别命名为BvACT1和BvACT2,已在GenBank中注册,登录号为KF214784和KF214785。多重序列比较表明,Bv A CT1氨基酸序列与其他植物Actin基因的同源性在92%以上,而Bv A CT2则在93%以上。

Cloning and Sequence Analysis of Actin Gene Fragments from Sugarbeet

In this study, a pair of degenerate primers were designed based on the conserved sequences of Actin genes from the other plants, and total RNA in roots of sugarbeet was used as template. The fragments of Actin gene were obtained by the RT-PCR method, then cloned into pUCm-T Vector. The positive clones were identified through the PCR method, and sequenced by Shanghai Sangon. Analysis of sequences showed that two members of Actin gene family were obtained, whose fragments length were both 598 bp, encoding 198 amino acids, and identities of 87% among them. They were named as BvA CT1 and BvA CT2, and deposited at GenBank under accessions numbers KF214784 and KF214785, respectively. Multiple sequence alignment revealed that amino acid sequences of Bv A CT1 and Bv A CT2 identities to the other plants Actin genes were above 92% and 93%, respectively.