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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (13): 243-248.doi: 10.11924/j.issn.1000-6850.2011-3582

所属专题： 生物技术

• 林学 园艺 园林 • 上一篇    下一篇

萼脊兰Actin基因片段的克隆及序列分析

袁秀云 田云芳 蒋素华 崔波   

  • 收稿日期:2011-12-01 修回日期:2012-02-01 出版日期:2012-05-05 发布日期:2012-05-05
  • 基金资助:

    郑州市科技计划项目

Cloning and Sequence Analysis of Actin Gene Fragment from Sedirea japonica

  • Received:2011-12-01 Revised:2012-02-01 Online:2012-05-05 Published:2012-05-05

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摘要：

从萼脊兰叶片中克隆Actin基因，为研究其生理功能及其他基因在萼脊兰中的表达和调控奠定基础。根据GenBank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，以萼脊兰叶片总RNA为模板，利用RT-PCR技术得到了3个Actin基因片段。序列分析结果表明，3个Actin基因片段长度均为1062 bp，编码354个氨基酸，具有Actin基因的功能位点；与其他植物同源序列进行分析表明，其核苷酸序列的同源性在80%以上，氨基酸序列的同源性在96%以上。本研究中得到的3个基因序列是Actin基因的同源片段，分别命名为SeACT1、SeACT2和SeACT3，并在GenBank注册，登录号分别为JN981138、JN981139和JN981140。

关键词: 品质比较, 品质比较

Abstract:

The aim was to clone Actin gene from leaf of Sedirea japonica, the results would provide an important basis to study the function of Actin and to research the molecular regulatory mechanisms of other genes as a reference gene. Degenerate primers were designed based on the conserved sequences of the Actin genes from other plants. Total RNA was extracted from the leaves of Sedirea japonica. Three Actin gene fragments were obtained by reverse transcription polymerase chain reaction (RT-PCR). The sequencing result revealed that each of three Actin gene fragments from Sedirea japonica contains 1062 bp, encoding a protein of 354 amino acids. Homology comparison with other plants Actin gene sequences in the GenBank showed that they shared over 80% nucleotide sequence homology and 96% amino acid sequence homology with other plants Actins. The cloned genes were Actin gene fragments, were named as SeACT1, SeACT2 and SeACT3 respectively. They were registered into GenBank (respective accession number: JN981138, JN981139 and JN981140).