大豆C2H2型锌指蛋白基因SCTF-1的克隆及功能分析

宋冰，王丕武，付永平，范旭红，夏海丰，高玮，洪洋，王贺，张卓，马建

doi:10.3724/SP.J.1005.2012.00749

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2012 , Vol. 34 >Issue 6: 749 - 756

DOI: https://doi.org/10.3724/SP.J.1005.2012.00749

研究报告

大豆C₂H₂型锌指蛋白基因SCTF-1的克隆及功能分析

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1. 吉林农业大学农学院, 长春 130118 2. 吉林市农业科学院, 吉林市132101 3. 吉林省农业科学院大豆研究中心, 长春130033 4. 通化市农业科学院, 通化 135000

收稿日期: 2012-02-09

修回日期: 2012-03-02

网络出版日期: 2012-06-25

基金资助

国家自然科学基金项目(编号：30971805, 31171568), 教育部博士点基金项目(编号：20070193005),吉林省科学技术厅项目(编号：20090203), 吉林省发改委项目(编号：203-421)和长春市科技局项目(编号：09KZ26)资助

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Cloning and functional analysis of SCTF-1 encoding a C₂H₂-type zinc finger protein from soybean

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1. College of Agriculture, Jilin Agricultural University, Changchun 130118, China 2. Jilin City Academy of Agricultural Sciences, Jilin City 132101, China 3. Soybean Research Center, Jilin Academy of Agricultural Sciences, Changchun 130033, China 4. Tonghua Academy of Agricultural Sciences, Tonghua 135000, China

Received date: 2012-02-09

Revised date: 2012-03-02

Online published: 2012-06-25

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摘要

锌指蛋白是一类具有手指型结构的蛋白质, 其中一些锌指蛋白是转录因子, 对真核生物的生长发育及非生物逆境胁迫的耐受能力都有着重要作用。文章从大豆(Glycine max (L.) Merr.)中克隆了一个新的C₂H₂型锌指蛋白基因SCTF-1 (GenBank登录号：JQ692081), 该基因包含一个699 bp的开放阅读框, 编码233个氨基酸, 无内含子, 有两个典型的C₂H₂型锌指结构。锌指结构中有植物锌指蛋白特有的保守氨基酸序列QALGGH。经软件预测分析, 其等电点pI=8.33, 分子量24.9 kDa。农杆菌介导的洋葱表皮细胞GFP瞬时表达实验结果表明, SCTF-1蛋白能够定位到细胞核中。通过RT-PCR检测发现该基因在大豆叶和花中的表达量较高, 在茎和根的表达量相对较低。在对大豆幼苗的低温胁迫中, SCTF-1基因的表达量明显增加。将SCTF-1基因转入烟草(Nicotiana ta-bacum L.)中, 发现SCTF-1基因的过量表达能够明显提高转基因烟草的耐冷能力。

关键词： 大豆; 锌指蛋白; C₂H₂型; SCTF-1基因; 亚细胞定位; 耐冷能力

本文引用格式

宋冰，王丕武，付永平，范旭红，夏海丰，高玮，洪洋，王贺，张卓，马建 . 大豆C₂H₂型锌指蛋白基因SCTF-1的克隆及功能分析[J]. 遗传, 2012 , 34(6) : 749 -756 . DOI: 10.3724/SP.J.1005.2012.00749

Abstract

The zinc finger protein is one of the proteins with finger-like domain. Some of them are transcription factors which play important role in plant growth and plant resistance to abiotic stresses. In this paper, a novel C₂H₂-type zinc finger protein gene SCTF-1 (GenBank accession number JQ692081) was isolated from soybean (Glycine max (L.) Merr.) This gene has a 699 bp ORF (open reading frame) with no in-tron and encodes a 24.9 kDa protein with 233 amino acids. Its isoelectric point (pI) is 8.33. The SCTF-1 protein contains two typical C₂H₂-type zinc finger domains. Both of them have highly conserved amino acid sequence-QALGGH which is a particular characteristic of plant. Transient expression of the GFP-SCTF-1 protein in onion epidermal cell showed that SCTF-1 was localized in cell nuclei. RT-PCR results showed that SCTF-1 gene was expressed with high levels in flowers and leaves in soybean, but low in roots and stems. The expression of SCTF-1 gene was strongly induced by low temperature in the soybean seedlings. Overexpression of SCTF-1 enhanced cold tolerance of transgenic tobacco (Nicotiana tabacum L.) compared to the control.

Key words： soybean; zinc finger protein; C₂H₂-type; SCTF-1 gene; subcellular localization; cold tolerance

参考文献

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