| 荔波连蕊茶肌动蛋白基因全长cDNA的克隆及其表达分析 |
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| 引用本文： | 肖政,李纪元,范正琪,殷恒福.荔波连蕊茶肌动蛋白基因全长cDNA的克隆及其表达分析[J].林业科学研究,2014,27(3):374-380. |
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| 作者姓名： | 肖政 李纪元 范正琪 殷恒福 |
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| 作者单位： | 中国林业科学研究院亚热带林业研究所, 浙江 富阳 311400;中国林业科学研究院亚热带林业研究所, 浙江 富阳 311400;中国林业科学研究院亚热带林业研究所, 浙江 富阳 311400;中国林业科学研究院亚热带林业研究所, 浙江 富阳 311400 |
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| 基金项目： | 国家“十二五”科技支撑计划课题（2012BA01B0703）；国家国际科技合作项目（2011DFA30490）；浙江省花卉新品种选育重大科技专项（2012C12909-6）；浙江省省院合作林业科技项目（2012SY02） |
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| 摘 要： | 根据植物肌动蛋白(Actin)基因编码区的保守序列设计引物,以山茶属植物荔波连蕊茶幼嫩茎段为材料,提取总RNA,进行RT-PCR。采用RACE技术扩增获得1 631 bp的Actin基因全长cDNA序列,命名为ClActin1(GenBank登录号KF366912)。序列分析表明,ClActin1开放阅读框(ORF)为1 134 bp,编码377个氨基酸,5’非编码区90 bp,3’非编码区407 bp。预测的ClActin1蛋白分子量为41.69 kD,理论等电点为5.31,具有Actin超基因家族的保守结构域和肌动蛋白家族特有的特征信号序列。ClActin1与GenBank中收录的其它植物肌动蛋白核苷酸序列的相似性在82%以上,氨基酸序列的相似性在97%以上。与其它植物肌动蛋白比较并构建系统进化树,结果显示山茶肌动蛋白与茶树和杨树的肌动蛋白的亲缘关系最为密切。实时荧光定量PCR结果显示,该基因在荔波边蕊茶不同组织器官及不同发育时期的表达量稳定,表明ClActin1基因可作为内参基因。
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| 关 键 词： | 荔波连蕊茶 肌动蛋白基因 序列分析 克隆 |
| 收稿时间： | 2013/7/17 0:00:00 |
Cloning and Expression Analysis of Full Length cDNA of Actin Gene from Camellia lipoensis |
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| XIAO Zheng,LI Ji-yuan,FAN Zheng-qi and YIN Heng-fu.Cloning and Expression Analysis of Full Length cDNA of Actin Gene from Camellia lipoensis[J].Forest Research,2014,27(3):374-380. |
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| Authors: | XIAO Zheng LI Ji-yuan FAN Zheng-qi and YIN Heng-fu |
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| Institution: | Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Fuyang 311400, Zhejiang, China;Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Fuyang 311400, Zhejiang, China;Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Fuyang 311400, Zhejiang, China;Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Fuyang 311400, Zhejiang, China |
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| Abstract: | In this study, the actin gene was cloned from the stems of Camellia lipoensis using RT-PCR and RACE methods. The full length of the actin gene, named ClActin1, was 1 631 bp (GenBank accession No.KF366912), which contains a 1 134 bp open reading frame (ORF) encoding 377 amino acid residues, contains a 5'-UTR with 90 bp and 3'-UTR with 407 bp. The putative protein molecular weight is 41.69 kD and its theoretical isoelectric point is 5.31. The sequence includes actin superfamily domain and the characteristic actin family signature sequences. Homologous alignment shows that it shares over 82% of nucleotide identities and over 97% of amino acid identities with actins from other plants in GenBank. The phylogenetic tree constructed on the basis of amino acid sequences suggests that the relationship of actins from C. lipoensis is most intimate with that from C. sinensis and Populus trichocarpa. The results from the analysis on tissue specific expression show that the endogenous actin gene expression level in the root, stem, leaf and seed of C. lipoensis is identical. It can be served as inner control to determine the relative expression amount of target genes from Camellia plants. |
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| Keywords: | Camellia lipoensis actin gene sequence analysis cloning |
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