南方农业学报2017，Vol.48Issue(5)：761-767,7.DOI:10.3969/j.issn.2095-1191.2017.05.001
二穗短柄草钙依赖型蛋白激酶基因 BdCDPK14克隆及表达分析
Clone and expression of calcium-dependent protein kinase BdCDPK14 gene from Brachypodium distachyon
摘要
Abstract
[Objective]The present experiment was conducted to clone calcium dependent protein kinase (CDPK) Bd-CDPK14 gene from Brachypodium distachyon and detect its expression level under drought stress, in order to provide a foundation for further research on regulation mechanism of BdCDPK14 gene in response to drought. [Method]Specific primers were designed based on NCBI retrieval results, and cDNA of B. distachyon was used as template. BdCDPK14, one of B. distachyon CDPK genes was cloned by PCR amplification. Encoded protein of BdCDPK14 gene was analyzed by online software on its bioinformatics. Its expression level under PEG-6000 drought stress was detected by RT-PCR. [Result]Bd-CDPK14 gene(GenBank accession number: XM_003564390) cloned from B. distachyon was 1750 bp in length, open reading frame(ORF) was 1545 bp CDS, coding 514 amino acids. The molecular weight of encoded protein was 56.78 kD, isoelectric point 5.45, fat index 78.21 and instability index 38.66, belonging to stable protein. BdCDPK14 protein con-tained several domains including four EF-hands, protein tyrosine kinase domain, lipopolysaccharide kinase family, ATP-binding region, serine/threonine protein kinase active-site region and predicted transmembrane region. It shared the closest genetic relationship with TaCPK13 protain(ABY59018) of wheat. BdCDPK14 protein was a protein with random coil andα-helix located in chloroplast and plasma membrane. [Conclusion]The cloned BdCDPK14 gene from B. distachyon leaf is a member of CDPK genes, and is involved in drought-resistant reaction in B. distachyon. So it can be served as a candi-date gene to further study its role in drought-resistant in B. distachyon.关键词
二穗短柄草/BdCDPK14基因/克隆/生物信息学分析/干旱胁迫/表达Key words
Brachypodium distachyon/BdCDPK14 gene/clone/bioinformatics analysis/drought stress/expression分类
农业科技引用本文复制引用
韦淑亚,赵旭东,杨广笑,何光源..二穗短柄草钙依赖型蛋白激酶基因 BdCDPK14克隆及表达分析[J].南方农业学报,2017,48(5):761-767,7.基金项目
湖北省自然科学研究计划项目(B2016581) （B2016581）
教育部高校博士学科点专项科研基金项目(20120142110075) （20120142110075）
湖北省高等学校优秀中青年科技创新团队计划项目(T201535) （T201535）
武汉职业技术学院博士科研基金项目(2016BS001) （2016BS001）
