大豆C2H2型锌指蛋白基因SCTF-1的克隆及功能分析

doi:10.3724/SP.J.1005.2012.00749

遗传 ›› 2012, Vol. 34 ›› Issue (6): 749-756.doi: 10.3724/SP.J.1005.2012.00749

大豆C₂H₂型锌指蛋白基因SCTF-1的克隆及功能分析

宋冰^1,2, 王丕武¹, 付永平¹, 范旭红³, 夏海丰^1,4, 高玮⁴, 洪洋¹, 王贺¹, 张卓¹, 马建¹

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 出版日期:2012-06-20 发布日期:2012-06-25
通讯作者: 王丕武 E-mail:peiwuw@yahoo.com.cn
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

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

SONG Bing^1,2, WANG Pi-Wu¹, FU Yong-Ping¹, FAN Xu-Hong³, XIA Hai-Feng^1,4, GAO-Wei⁴, HONG Yang¹, WANG He¹, ZHANG Zhuo¹, MA Jian¹

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:2012-02-09 Revised:2012-03-02 Online:2012-06-20 Published:2012-06-25

摘要/Abstract

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

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

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.

SONG Bing, WANG Pi-Wu, FU Yong-Ping, FAN Xu-Hong, XIA Hai-Feng, GAO-Wei, HONG Yang, WANG He, ZHANG Zhuo, MA Jian. Cloning and functional analysis of SCTF-1 encoding a C₂H₂-type zinc finger protein from soybean[J]. HEREDITAS(Beijing), 2012, 34(6): 749-756.

参考文献

[1] Sakamoto H, Araki T, Meshi T, Iwabuchi M. Expression of a subset of the Arabidopsis Cys₂/His₂-type zinc-finger protein gene family under water stress. Gene, 2000, 248(1-2): 23-32.
[2] Sugano S, Kaminaka H, Rybka Z, Catala R, Salinas J, Matsui K, Ohme-Takagi M, Takatsuji H. Stress-responsive zinc finger gene ZPT2-3 plays a role in drought tolerance in petunia. Plant J, 2003, 36(6): 830-841.
[3] 郭书巧, 黄骥, 江燕, 张红生. 水稻C2H2型锌指蛋白基因RZF71的克隆与表达分析. 遗传, 2007, 29(5): 607-618.
[4] 郭书巧, 张红生. 水稻锌指蛋白基因RZF5和RZF71的克隆与功能分析
[学位论文]. 南京: 南京农业大学, 2006: 23-120.
[5] Kim JC, Lee SH, Cheong YH, Yoo CM, Lee SI, Chun HJ, Yun DJ, Hong JC, Lee SY, Lim CO, Cho MJ. A novel cold-inducible zinc finger protein from soybean, SCOF-1, enhances cold tolerance in transgenic plants. Plant J, 2001, 25(3): 247-259.
[6] 邢浩然, 刘丽娟, 刘国振. 植物蛋白质的亚细胞定位研究进展. 华北农学报, 2006, 21(增刊): 1-6.
[7] 黄骥, 王建飞, 张红生. 植物C2H2 型锌指蛋白的结构与功能. 遗传, 2004, 26(3): 414-418.
[8] 刘海燕, 冯冬茹, 刘兵, 何炎明, 王宏斌, 王金发. 农杆菌介导的MpASR蛋白在洋葱表皮细胞的定位研究. 热带亚热带植物学报, 2009, 17(3): 218-222.
[9] 黄骥, 张红生. TFⅢA型锌指蛋白及在提高植物耐逆性中的作用. 遗传, 2007, 29(8): 915-922.
[10] Dinkins RD, Pflipsen C, Collins GB. Expression and deletion analysis of an Arabidopsis SUPERMAN-like zinc finger gene. Plant Sci, 2003, 165(1): 33-41.
[11] Payne T, Johnson SD, Koltunow AM. KNUCKLES (KNU) encodes a C2H2 zinc-finger protein that regulates devel-opment of basal pattern elements of the Arabidopsis gy-noecium. Development, 2004, 131(15): 3737-3749.
[12] Nakagawa H, Jiang CJ, Sakakibara H, Kojima M, Honda I, Ajisaka H, Nishijima T, Koshioka M, Homma T, Mander LN, Takatsuji H. Overexpression of a petunia zinc-finger gene alters cytokinin metabolism and plant forms. Plant J, 2005, 41(4): 512-521.
[13] Huang F, Chi YJ, Meng QC, Gai JY, Yu DY. GmZFP1 encoding a single zinc finger protein is expressed with enhancement in reproductive organs and late seed development in soybean (Glycine max). Mol Biol Rep, 2006, 33(4): 279-285.
[14] Lippuner V, Cyert MS, Gasser CS. Two classes of plant cDNA clones differentially complement yeast calcineurin mutants and increase salt tolerance of wild-type yeast. J Biol Chem, 1996, 271(22): 12859-12866.
[15] Sakamoto H, Araki T, Meshi T, Iwabuchi M. Expression of a subset of the Arabidopsis Cys₂/His₂-type zinc-finger protein gene family under water stress. Gene, 2000, 248(1-2): 23-31.
[16] van Der Krol AR, van Poecke RM, Vorst OF, Voogt C, van Leeuwen W, Borst-Vrensen TW, Takatsuji H, van Der Plas LH. Developmental and wound-, cold-, desiccation-, ul-traviolet-B-stress-induced modulations in the expression of the petunia zinc finger transcription factor gene ZPT2-2. Plant Physiol, 1999, 121(4): 1153-1162.
[17] 白晶, 张必弦, 李新玲, 徐香玲. 野生大豆(Glycine soja) C2H2型锌指蛋白基因的克隆与序列分析. 大豆科学, 2009, 28(1): 21-25.

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大豆C₂H₂型锌指蛋白基因SCTF-1的克隆及功能分析

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

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