摘要
以文心兰叶片为材料,根据Gen Bank中登录的植物肌动蛋白基因同源核苷酸保守序列设计简并引物,利用RT-PCR技术得到了2个动蛋白基因(Actin)片段。序列分析结果表明,2个Actin基因片段长度均为1062 bp,2条序列一致性为90.87%,分别命名为OnA CT1和OnA CT2,并在Gen Bank注册,登录号分别为JN981136和JN981137。2条序列编码完全相同的354个氨基酸,具有Actin蛋白的特征序列;进化树分析表明,文心兰Actin蛋白与萼脊兰和蕙兰的亲缘关系最近。采用实时荧光定量(qR T-PCR)和半定量RT-PCR(Semi-quantitative RT-PCR)方法进行表达分析,结果显示该基因在营养生长和生殖生长阶段的不同组织中表达相对稳定,可以作为发育阶段相关基因表达研究的内参基因。
The aim of this study was to clone the Actin gene from the leaf of Oncidium ＇Sweet Sugar＇. De- generate primers were designed based on the registered sequences of the Actin genes from other plants. Two Actin sequence fragments were obtained by RT-PCR. The sequencing result revealed that the two Actin gene fragments from Oncidium ＇Sweet Sugar＇ contain 1062 bp with 90.87% homology, which were named as OnACT1 and OnACT2 and registered into GenBank （respective accession number： JN981136 and JN981137）. They encode the same protein of 354 amino acids with three Actin signatures. Phylogenetic tree analysis indicated that Actin from Oncidium＇Sweet Sugar was closely related to the A ctin of Sedireajaponica and Cymbidiitm faberi. The expression ofActin gene was analyzed using qRT-PCR and semi-quantitative RT-PCR. The result showed that the expression of Actin was relatively stable in different tissues at the vegetative and reproductive stage; therefore, it could be used as reference genes of the development related genes.
出处
《安徽农业大学学报》
CAS
CSCD
北大核心
2016年第1期77-82,共6页
Journal of Anhui Agricultural University
关键词
ACTIN基因
克隆
序列分析
文心兰
Actin gene
cloning
sequence analysis
Oncidium ＇Sweet Sugar＇
作者简介
邓祖丽颖，副教授。E—mail：13938289841@139．com
