Paper - WANFANG MED ONLINE

You Position: Home > Paper

Cloning of Copper Chaperone for Superoxide Dismutase Gene CCS from Embryogenic Callus of Dimocarpus longan Lour.and Its Expression Analysis During Somatic Embryogenesis

( views:69, downloads:0 )

Author:: No author available

Journal Title:: Chinese Journal of Applied & Environmental Biology

Issue:: 3

DOI:: 10.3724/SP.J.1145.2012.00351

Key Word:: 龙眼;体细胞胚胎发生;CCS;克隆;生物信息学;实时荧光定量PCR

Abstract：为了解Cu/Zn超氧化物歧化酶分子伴侣基因(CCS)在龙眼体胚发生过程中的表达调控机制,以龙眼松散型胚性愈伤组织为材料,采用RT-PCR法,克隆了长960 bp含有完整开放阅读框的龙眼DlCCS核酸序列,其编码一个含有319个氨基酸的蛋白质(GenBank登录号:FJ973472).生物信息学分析显示:该蛋白为亲水的、稳定的、含有跨膜结构域的酸性蛋白质,定位于叶绿体；与毛果杨、葡萄、拟南芥、水稻和锦鸡儿具有较高同源性,含有植物CCS蛋白所特有的3个保守结构域；预测其通过不同的磷酸化方式,改变酶活性及蛋白构象,参与龙眼体胚中超氧化物代谢以及金属离子转运等生物学过程,在氧化还原过程中发挥作用.利用实时荧光定量PCR (Qpcr)技术研究该基因在龙眼体胚发生发育过程中的表达情况,结果显示,从松散型胚性愈伤组织( Stage 1)到胚性紧实球形结构阶段(Stage 4),DlCCS Mrna的转录水平较低且波动小；当胚性紧实球形结构进一步发育到子叶形胚阶段(Stage 8),其表达量迅速增加,在成熟胚阶段( Stage 9)达到最高峰,提示DlCCS可能在龙眼体胚的中晚期起主要作用.

Reference

[1]张巧,汪静儿,叶春丽,等.四倍体野生棉毛棉体细胞胚胎发生与植株再生研究[J].2010,(3).
[2]姜铮,王芳,何湘,等.蛋白质磷酸化修饰的研究进展[J].2009,(2).
[3]赖钟雄,陈春玲.龙眼体细胞胚胎发生过程中的内源激素变化[J].2002,(2).
[4]Yick PJ,Culotta VC,Leitch JM.The right to choose: multiple pathways for activating copper, zinc superoxide dismutase.[J].2009,284(37).
[5]赖呈纯,赖钟雄,方智振,等.龙眼胚性愈伤组织线粒体ATP合酶β亚基基因克隆及其在龙眼体胚发生过程中的表达分析[J].2010,(16).
[6]Trindade LM,Visser RGF,Horvath BM,etc.Isolation of a gene encoding a copper chaperone for the copper/zinc superoxide dismutase and characterization of its promoter in potato[J].2003,133(2).
[7]Yu, Agnc.s,Bouchcb, Nicolas,Beauclair, Linda.microRNA-directed cleavage and translational repression of the copper chaperone for superoxide dismutase mRNA in Arabidopsis[J].2010,62(3).
[8]Wintz H.,Vulpe C..Plant copper chaperones[J].2002.
[9]Schmidt PJ.,Culotta VC.,Kunst C..Copper activation of superoxide dismutase 1 (SOD1) in vivo - Role for protein-protein interactions with the copper chaperone for SOD1[J].2000,275(43).
[10]A L, Lamb,A S, Torres,T V, O'Halloran,etc.Heterodimeric structure of superoxide dismutase in complex with its metallochaperone.[J].2001,8(9).
[11]Lai Z.X.,He Y.,Chen Y.T.,etc.Molecular Biology and Proteomics During Somatic Embryogenesis in Dimocarpus Longan Lour.[J].2010.
[12]J M, McCord,I, Fridovich.Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein).[J].1969,244(22).
[13]Lin, Y.L.,Lai, Z.X..Reference gene selection for qPCR analysis during somatic embryogenesis in longan tree[J].2010,178(4).
[14]Culotta VC,Pufahl RA,Schmidt PJ,etc.Multiple protein domains contribute to the action of the copper chaperone for superoxide dismutase.[J].1999,274(34).
[15]Pullman GS.,Cairney J.,Xu NF.,etc.Loblolly pine (Pinus taeda L.) somatic embryogenesis: maturation improvements by metal analyses of zygotic and somatic embryos[J].2003,164(6).
[16]Leitch, Jeifry M.,Jensen, Laran T.,BouIdin, Samantha D.,etc.Activation of Cu, Zn-Superoxide Dismutase in the Absence of Oxygen and the Copper Chaperone CCS.[J].2009,284(33).