Cloning and Bioinformatics Analysis of a DsMAPKKK Gene from Dunaliella salina

Chinese Agricultural Science Bulletin ›› 2014, Vol. 30 ›› Issue (15) : 274-281. DOI: 10.11924/j.issn.1000-6850.2013-2740
23

Cloning and Bioinformatics Analysis of a DsMAPKKK Gene from Dunaliella salina

Author information +
History +

Abstract

To illustrat the function of MAPKs cascade reaction in salt-tolerance mechanism of Dunaliella salina, it was necessary to clone a MAPKKK gene which was a member of MAPKs signal transduction pathway and study its function.A mitogen-activated protein kinase kinase kinase gene was firstly cloned from Dunaliella salina by RT-PCR and RACE technology and named as DsMAPKKK(GenBank Accession No.KF366904) and further bioinformatics-analysis was conducted.The result showed that, full length of cDNA for DsMAPKKK gene was 1460 bp with a 879-bp open reading frame coding 292 amino acids.This protein was unstable, hydrophilic and located in cytoplasmic matrix without signal peptide and transmembrane region.The protein sequence contained a protein kinases ATP-binding region spanning from 24rd to 45rd amino acid and a Serine/Threonine protein kinase region spanning from 137rd to 149rd amino acid.Several potential phosphorylation sites and an important catalytic active site Asp141 were found in this protein.The main components of protein secondary structure were α-helix and random coil.A 3D model of protein tertiary structure was built successfully.Phylogenic analysis showed this protein had the closest evolutionary relationship with the corresponding proteins of Volvox carteri f. Nagariensis and Chlamydomonas reinhardtii.

Key words

Dunaliella salina; MAPKKK; full length of cDNA; bioinformatics

Cite this article

Download Citations
Cloning and Bioinformatics Analysis of a DsMAPKKK Gene from Dunaliella salina. Chinese Agricultural Science Bulletin. 2014, 30(15): 274-281 https://doi.org/10.11924/j.issn.1000-6850.2013-2740

References

[1] 孟祥宗.盐藻功能基因组学研究—研究资源的构建及重要基因的分离和鉴定[D].上海:上海大学,2006:133-138.
[2] 张婷,柴晓杰.王媛,等.盐生杜氏藻MAPK基因的克隆及表达分析[J].中国农学通报,2013,29(15):157-163.
[3] Mizoguchia T, Iriet K, Hirayama T, et al. A gene encoding a mitogen- activated protein kinase kinase kinase is induced simultaneously with genes for a mitogen- activated protein kinase and an S6 ribosomal protein kinase by touch, cold, and water stress in Arabidopsis thaliana[J]. Plant Biology,1996,93:765-766.
[4] Ichimura K, Shinozaki G, Sheen J, et al. Mitogen-activated protein kinase cascades in plants:a new nomenclature[J].Plant Science,2002, 7(7):306-307.
[5] 赵丽娜,陈喜文,陈德富.甘油代谢对杜氏盐藻盐耐受性的调节[J].生物技术通讯,2011,22(3):428-432.
[6] 陈志,焦新之,刘虹.杜氏盐藻质膜AT Pase在渗透调节中的可能作用[J].植物生理学报,1991,17(4):333-341.
[7] Alkayala F, Albionb R, Tillett R, et al. Expressed sequence tag (EST) profiling in hyper saline shocked Dunaliella salina reveals high expression of protein synthetic apparatus components [J]. Plant Sci, 2010, 179(5): 437-449.
[8] Zhang X, Qu Z, Wan Y, et al. Application of suppression subtractive hybridization (SSH) to cloning differentially expreeed cDNA in Dunaliella salina (Chlorophyta) under hyperosmotic shock [J]. Plant Mol Biol Rep, 2002, 20:49-57.
[9] 张晓琳,柴晓杰,张婷,等.盐藻 CDPK 基因的克隆与生物信息学分析[J].核农学报,2013,27( 4) :0418-0420.
[10] 汪虹,鲍大鹏,陈明杰,等.草菇MAPKKK 同源基因的克隆及序列分析[J].微生物学通报,2010,37(11):1606-1609.
[11] 石林丹.鉴定导致程序性死亡的线性级联信号通路中的三个MAPKKKs[J].农业生物技术学报,2012,20(7):755-760.
[12] 韩静.水稻MAPKK家族基因克隆及转基因研究[D].上海:上海海洋大学,2009:9-16.
[13] Gao L, Xiang C B. The genetic locus At1g73660 encodes a putative MAPKKK and negatively regulates salt tolerance in Arabidopsis[J]. Plant Mol Biol,2008,67(1-2):125-34.
[14] Yin Z J, Wang J J, Wang D L, et al. The MAPKKK Gene Family in Gossypium raimondii: Genome- Wide Identification, Classification and Expression Analysis[J].Int. J. Mol. Sci,2013,14:18740-18760.
[15] 薛飞,柴晓杰,余祝君,等.盐藻新基因DsSTPK的克隆及生物信息学分析[J].中国生物化学与分子生物学报,2012,28(3):88-91.
[16] 刘春燕,魏学军,张娜,等.TcLr45凝集素类受体蛋白激酶基因克隆与序列分析[J].植物遗传资源学报2013,14(3):489-491.
[17] 周俊琴,谭晓风,袁军,等.油茶Pht1;2的克隆及生物信息学分析[J].生物技术,2013,23(1):38-40.
[18] Sinha A K, Jaggi M, Raghuram B, et al. Mitogen-activated protein kinase signaling in plants under abiotic stress[J]. Plant Signaling & Behavior, 2011,6(2):196-203.
[19] Sabeeha S, Simon E, Olivier V, et al. The Chlamydomonas Genome Reveals the Evolution of Key Animal and Plant Functions[J]. Science,2007,318(5848):245-250.
Share on Mendeley

Accesses

Citation

Detail

Sections
Recommended

/