二穗短柄草钙依赖型蛋白激酶BdCDPK4基因的克隆及其编码蛋白质的生物信息学分析

doi:10.3969/j.issn.1004-1524.2017.02.23

浙江农业学报 ›› 2017, Vol. 29 ›› Issue (2): 345-352.DOI: 10.3969/j.issn.1004-1524.2017.02.23

• 生物系统工程 • 上一篇

二穗短柄草钙依赖型蛋白激酶BdCDPK4基因的克隆及其编码蛋白质的生物信息学分析

韦淑亚¹, 赵旭东², 王会平¹, 杨广笑², 何光源²

1.武汉职业技术学院生物工程学院应用生物技术研究所,湖北武汉 430074;
2. 科技部国际科技合作基地(基因工程),分子生物物理学教育部重点实验室,华中科技大学生命科学与技术学院,湖北武汉 430074

收稿日期:2016-10-01 出版日期:2017-02-15 发布日期:2017-03-06
作者简介:韦淑亚(1979—),女,湖北武汉人,博士,主要从事生物化学与分子生物学研究。E-mail:weishuya1011@126.com
基金资助:
高校博士点专项科研基金(20120142110075); 湖北省教育厅自然科学研究计划项目(B2016581); 河南省教育厅自然科学研究计划项目(2009B210005); 武汉职业技术学院校级博士科研基金(2016BS001)

Cloning and encoding protein bioinformatics analysis of BdCDPK4 from Brachypodium distachyon

WEI Shuya¹, ZHAO Xudong², WANG Huiping¹, YANG Guangxiao², HE Guangyuan²

1. Institute of Applied Bio-technology, College of Biological Engineering, Wuhan Polytechnic, Wuhan 430074, China;
2. The Genetic Engineering International Cooperation Base of Ministry of Science and Technology, Key Laboratory of Molecular Biophysics of Chinese Ministry of Education, College of Life Science and Technology, Huazhong University of Science & Technology, Wuhan 430074, China;

Received:2016-10-01 Online:2017-02-15 Published:2017-03-06

摘要/Abstract

摘要： 二穗短柄草(Brachypodium distachyon L.)因与小麦、大麦等有很高的同源性,作为新型禾本科模式生物吸引了越来越多的研究者。试验采用基因克隆技术分离了二穗短柄草BdCDPK4基因的编码区序列,并采用生物信息学方法分析了该基因及其编码的蛋白质的序列特征、结构和功能。结果表明,分离了BdCDPK4基因 1 851 bp 的 cDNA 序列(GenBank 登录号为 XM_003559516),编码区长度为 1 752 bp,编码 583 个氨基酸,分子量为 64.78 ku,等电点为9.12,含有N端可变区、蛋白激酶区、自抑制区、EF手型结构、类似钙调素等结构域。亚细胞定位预测位于叶绿体和液泡上。进化树分析结果显示,该蛋白与小麦TaCDPK19 蛋白的同源性最近。根据各物种间 CDPKs 蛋白的高同源性,结合小麦TaCDPK19基因被高盐和小麦白粉病菌Blumeria graministritici (Bgt)诱导,推测BdCDPK4基因也可能参与逆境胁迫下的信号传导途径,在抗逆等生物学过程中发挥重要功能。

关键词: CDPKs, BdCDPK4, 多序列比对, 生物信息学分析

Abstract: Brachypodium distachyon, the genome of which shares high homology with wheat and barley, has been used as the new model species of Gramineae. In order to clarify the structure and function of Brachypodium distachyon CDPKs gene, the coding region sequence (CDS) of BdCDPK4 gene of Brachypodium distachyon was isolated, and its character, structure and function were analyzed by bioinformatics methods. The results showed that 1 851 bp cDNA (GenBank ID: XM_003559516) of coding sequence of BdCDPK4 gene included the complete 1 752 bp CDS, coded 583 amino acids, and located on chloroplast and vacuole. The molecular weight and isoelectric point of BdCDPK4 protein were 64.78 ku and 9.12, respectively. BdCDPK4 protein contained several domains including N-variable domain, kinase domain, autoinhibitory domain, EF-hands and CaM-like domain. Phylogenetic analysis results suggested that BdCDPK4 protein had high homology with wheat TaCDPK19. TaCDPK19 were induced by high salt and white powder Blumeria graministritici(Bgt). BdCDPK4 protein may participate in the regulation of signal transduction pathways under stresses according to the homologies of protein domain among all species. Taken together, our data could establish a good foundation for studying the biological functions of BdCDPK4 gene.

Key words: CDPKs, BdCDPK4, multiple sequence alignment, bioinformatics analysis

中图分类号:

Q949.71⁺4.2

韦淑亚, 赵旭东, 王会平, 杨广笑, 何光源. 二穗短柄草钙依赖型蛋白激酶BdCDPK4基因的克隆及其编码蛋白质的生物信息学分析[J]. 浙江农业学报, 2017, 29(2): 345-352.

WEI Shuya, ZHAO Xudong, WANG Huiping, YANG Guangxiao, HE Guangyuan. Cloning and encoding protein bioinformatics analysis of BdCDPK4 from Brachypodium distachyon[J]. , 2017, 29(2): 345-352.

参考文献

[1] HETHERINGTON A, TREWAVAS A. Calcium-dependent protein kinase in pea shoot membranes[J]. FEBS Letters, 1982, 145 (1): 67-71.
[2] HARMON A C, PUTNAM-EVANS C, CORMIER M J. A calcium-dependent but calmodulin-independent protein kinase from soybean[J]. Plant Physiology, 1987, 83 (4): 830-837.
[3] CHENG S H, WILLMANN M R, CHEN H C, et al. Calcium signaling through protein kinases. The Arabidopsis calcium-dependent protein kinase gene family[J]. Plant Physiology, 2002, 129 (2): 469-485.
[4] ASANO T, TANAKA N, YANG G, et al. Genome-wide identification of the rice calcium-dependent protein kinase and its closely related kinase gene families: comprehensive analysis of the CDPKs gene family in rice[J]. Plant and Cell Physiology, 2005, 46 (2): 356-366.
[5] RAY S, AGARWAL P, ARORA R, et al. Expression analysis of calcium-dependent protein kinase gene family during reproductive development and abiotic stress conditions in rice (Oryza sativa L. ssp. indica) [J]. Molecular Genetics and Genomics, 2007, 278 (5): 493-505.
[6] 张进, 李建波, 刘伯斌, 等. 杨树CDPK基因家族的表达分析及功能预测[J]. 林业科学研究, 2014, 27 (5): 604-611.
ZHANG J, LI J B, LIU B B, et al. Expression and functional analysis of CDPK gene family in populus[J]. Forest Research, 2014, 27 (5): 604-611.(in Chinese with English abstract)
[7] SHEEN J. Ca 2+ -dependent protein kinases and stress signal transduction in plants[J]. Science, 1996, 274 (5294): 1900-1902.
[8] SAIJO Y, HATA S, KYOZUKA J, et al. Over-expression of a single Ca 2+ -dependent protein kinase confers both cold and salt/drought tolerance on rice plants[J]. Plant Journal, 2000, 23 (3): 319-327.
[9] SAIJO Y, KINOSHITA N, ISHIYAMA K, et al. A Ca 2+ -dependent protein kinase that endows rice plants with cold-and salt-stress tolerance functions in vascular bundles[J]. Plant and Cell Physiology, 2001, 42 (11): 1228-1233.
[10] WEI S, HU W, DENG X, et al. A rice calcium-dependent protein kinase OsCPK9 positively regulates drought stress tolerance and spikelet fertility[J]. BMC Plant Biology, 2014, 14 (1): 133.
[11] ZHU S Y, YU X C, WANG X J, et al. Two calcium-dependent protein kinases, CPK4 and CPK11, regulate abscisic acid signal transduction in Arabidopsis[J]. Plant Cell, 2007, 19 (10): 3019-3036.
[12] 杜利欣, 胡宗利, 张建苓, 等. 番茄 SlDP1 基因的克隆、生物信息学及表达特性分析[J]. 植物研究, 2014, 34(1):68-74.
DU L X, HU Z L, ZHANG J L, et al. Cloning, bioinformatics and expression analysis of SlDP1 gene in tomato[J]. Bulletin of Botanical Research, 2014, 34(1):68 -74. (in Chinese with English abstract)
[13] 梁志强, 孔德真, 李辉玲, 等. 小拟南芥 HDG12 基因的克隆、生物信息学分析及植物表达载体的构建[J]. 江苏农业科学, 2015, 43(2):33-37.
LIANG Z Q, KONG D Z, LI H L, et al. Cloning, bioinformatics and construction of plant expression vector of HDG12 in Arabidopsis pumila[J]. Jiangsu Agricultural Sciences, 2015, 43(2): 33-37.(in Chinese)
[14] LI A L, ZHU Y F, TAN X M, et al. Evolutionary and functional study of the CDPK gene family in wheat (Triticum aestivum L.) [J]. Plant Molecular Biology, 2008, 66(4):429-443.
[15] YU X, LI M, GAO G, et al. Abscisic acid stimulates a calcium-dependent protein kinase in grape berry[J]. Plant Physiology, 2006, 140(2):558-579.

二穗短柄草钙依赖型蛋白激酶BdCDPK4基因的克隆及其编码蛋白质的生物信息学分析

Cloning and encoding protein bioinformatics analysis of BdCDPK4 from Brachypodium distachyon

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 13

编辑推荐

Metrics

本文评价

[1]	赵秀平, 王双, 闫星伊, 段强, 张帅, 陈永胜, 李国瑞. 稻瘟病菌MGG-01005的表达纯化与生物信息学分析[J]. 浙江农业学报, 2021, 33(3): 470-478.
[2]	梁丽琴, 杨瑞, 郜刚. 马铃薯StUOXs基因家族的生物信息学分析[J]. 浙江农业学报, 2020, 32(9): 1523-1532.
[3]	吴佳, 陈朗, 姜涛, 黄国明, 李倬, 李耀东, 张丽, 刘丽霞. 奶牛CSF3基因遗传多态性筛查及其生物信息学分析[J]. 浙江农业学报, 2020, 32(6): 986-993.
[4]	刘正奎, 吴瑗, 陈琳, 王磊, 牟泓烨, 祝徐航, 王晓杜. 猪流行性腹泻病毒Nsp5基因的原核表达及生物信息学分析[J]. 浙江农业学报, 2019, 31(4): 532-538.
[5]	董新星, 李明丽, 崔艺佳, 兰国湘, 王孝义, 严达伟. 撒坝猪MCUR1基因克隆、生物信息学分析及其组织表达量检测[J]. 浙江农业学报, 2019, 31(11): 1825-1833.
[6]	李洪有, 霍冬敖, 蔡芳, 张晓娜, 石桃雄, 陈其皎, 陈庆富. 荞麦ARF基因家族的鉴定及生物信息学分析[J]. 浙江农业学报, 2018, 30(1): 7-7.
[7]	李文丽, 杜晓华, 刘霞, 高景波, 申超超, 张超, 刘伟. 藏绵羊FAM134B基因克隆与表达分析[J]. 浙江农业学报, 2017, 29(9): 1474-1481.
[8]	翟莹, 张军, 赵艳, 高士童, 孙婉姝, 张闯, 任巍巍, 王秀文, 王玉书. 大豆5个新发现ERF基因的生物信息学及表达分析[J]. 浙江农业学报, 2016, 28(10): 1644-1649.
[9]	赵莺婕1，刘春林2，阮颖1，*. 拟南芥AtSb10基因的克隆及其过表达转基因株系的获得[J]. 浙江农业学报, 2015, 27(9): 1550-.
[10]	潘蕾1，2，刘燕2，韦强2，肖琛闻2，季权安2，鲍国连2，，吴信生3，. 家兔MYL12B基因的克隆及生物信息学分析[J]. 浙江农业学报, 2015, 27(6): 927-.
[11]	方倩倩1，李祥龙2，*，周荣艳1，李兰会1. 不同物种Atrn基因编码区生物信息学分析[J]. 浙江农业学报, 2015, 27(6): 933-.
[12]	王同淑1，2，李传峰2，陈宗艳2，孟春春2，吴润1，，刘光清1，2，. 北京鸭RIG\\|I基因的克隆及其生物信息学分析 [J]. 浙江农业学报, 2015, 27(5): 734-.
[13]	彭静静. 植物中tRNase Z同源蛋白的鉴定和分析[J]. 浙江农业学报, 2014, 26(5): 1257-.