| 烟草钾转运体基因TPK1的电子克隆及生物信息学分析 |
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| 引用本文： | 鲁黎明.烟草钾转运体基因TPK1的电子克隆及生物信息学分析[J].中国农业科学,2011,44(1):28-35. |
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| 作者姓名： | 鲁黎明 |
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| 作者单位： | (四川农业大学农学院) |
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| 基金项目： | 国家植物生理生化重点实验室开放课题(PPB08008) |
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| 摘 要： | 目的]克隆烟草钾吸收转运基因,分析其亲缘关系,并预测其结构、性质与功能,为烟草钾转运体基因的功能研究提供基础.方法]以利用RACE技术克隆出的烟草钾转运体基因片段的3'端序列为探针,对烟草EST数据库进行搜索,筛选与探针同源性在95%以上的EST序列,再利用DNAStar软件进行拼接.然后,应用多种生物信息学数据库...
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| 关 键 词： | 烟草 钾转运体基因 生物信息学 Northern杂交 |
| 收稿时间： | 2010-06-11; |
In Silico Cloning and Bioinformatic Analysis of TPK1 Gene in Tobacco |
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| LU Li-ming.In Silico Cloning and Bioinformatic Analysis of TPK1 Gene in Tobacco[J].Scientia Agricultura Sinica,2011,44(1):28-35. |
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| Authors: | LU Li-ming |
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| Institution: | LU Li-ming1,2(1College of Agronomy,Sichuan Agricultural University,Chengdu 611134,2State Key Laboratory of Plant Physiology and Biochemistry,Beijing 100193) |
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| Abstract: | 【Objective】 The aim of this study is to clone K+ transporter gene in tobacco, analyze its genetic relationship, and predict its structure, nature, function and to provide a basis for K+ gene function studies of tobacco. 【Method】 Cloning tobacco K+ transporter gene by means of the combination of laboratory work and e-cloning techniques. Using tobacco K+ transporter gene 3' end sequence, cloned by RACE, as a probe to search the tobacco EST databases, filter out the ESTs sequences with the homology of 95% of the probe, and spliced the two segments using DNAStar software. Then, analyze gene structure and properties of the full cDNA sequence, and predict its function by applying a variety of bioinformatics databases and softwares. 【Result】A new tobacco K+ gene (TPK1) with full cDNA sequence was found. The molecular weight of the protein encoded by this gene is 26.21 kD, and the theoretical isoelectric point pI is 9.72. TPK1 is a hydrophobic membrane protein, containing six distinct transmembrane helix topologies. In secondary structure, a total of 11 α-helix, and 16 β-fold were found in TPK1. At subcellular level, TPK1 protein located in the plastid, vacuole and endoplasmic reticulum. The signal peptide splice site and the serine, threonine and tyrosine kinase phosphorylation sites in TPK1 amino acids sequence were found as well. In the function, TPK1 protein contains a typical conserved K+ uptake transporter function domain, and is highly of homology to the major K+ uptake transporter genes in higher plants. Northern blotting analysis indicated that TPK1 was expressed in root, stem, leaf and flower.【Conclusion】A new K+ transporter TPK1 was successfully cloned in tobacco, and its function maybe involved in K+ transportation in tobacco plants. Therefore, further investigation still needed to understand its role in tobacco K+ uptake and transportation. |
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| Keywords: | tobacco potassium transporter bioinfomatics Northern blotting |
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