全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

苋菜AmPCS基因的克隆及表达载体构建

DOI: 10.3969/j.issn.1000-7091.2012.02.009, PP. 44-49

Keywords: 苋菜,植物络合素,克隆,AmPCS基因,植物表达载体

Full-Text   Cite this paper   Add to My Lib

Abstract:

合成植物络合素(PCs)是高等植物减轻重金属毒害的重要机制。通过对镉超积累苋菜品种天星米植物络合素合成酶基因(PCS)的克隆及表达载体构建,旨在为植物修复镉污染土壤奠定基础。依据同源克隆原理,通过RT-PCR和RACE方法克隆苋菜PCS基因。序列分析表明:苋菜PCS基因cDNA全长1770bp,包含完整的阅读框,编码485个氨基酸;苋菜PCs合成酶与拟南芥、遏蓝菜、芥菜及油菜PCs合成酶相似性为91.96%~95.67%,其氨基酸序列N端有1个Pfam:Phytochelatin结构域(功能为催化合成植物络合素)、C端有1个Pfam:Phytochelatin_C结构域(功能为重金属离子感应器)。因此,推断苋菜PCS基因编码蛋白是PCS-like家族的新成员,具有催化合成植物络合素的功能,将它在GenBank注册,序列号为:JN979370,命名为AmPCS。在AmPCS基因的编码区加入BamHI和SacI酶切位点,双酶切植物表达载体pBI-121和带有酶切位点的PCR产物,成功获得苋菜PCS基因正义表达载体pBI-PCS。

References

[1]  Vatanmniuk O K,Mari S,Lu Y P,et al. AtPCS1,a phytochelatin synthase from Arabidopsis:isolation and in vitro reconstitution[J]. Proc Natl Acad Sci,1999,96:7l10-7l15.
[2]  Ha S B,Smith A P,Howden R,et al. Phytochelatin synthase genes from Arabidopsis and the yeast Schizosaccharomyces pombe[J]. Plant Cell,1999,11:l153- l164.
[3]  Huang H G,Li T X,Yang X E,et al. Research advances in?plant lead tolerance and detoxification mechanism[J]. Ying?Yong Sheng Tai Xue Bao,2009,20(3):696-704.
[4]  ?Liu GY,Zhang Y X,Chai TY. Phytochelatin synthase of?Thlaspi caerulescens enhanced tolerance and accumulation of?heavy metals when expressed in yeast and tobacco[J]. Plant?Cell Rep,2011,30(6):1067-1076.
[5]  Osaki Y,Shirabe T,Nakanishi H,et al. Characterization of?phytochelatin synthase produced by the primitive red alga?Cyanidioschyzon merolae[J]. Metallomics,2009,1(4):?353-358.
[6]  ?Wojas S,Clemens S,Sk?odowska A,et al. Arsenic response of AtPCS1-and CePCS-expressing plants -effects?of external As (V) concentration on As-accumulation?pattern and NPT metabolism[J]. J Plant Physiol,2010, 167(3):169-175.
[7]  Wojas S,Ruszczyńska A,Bulska E,et al. The role of subcellular distribution of cadmium and phytochelatins in the?generation of distinct phenotypes of AtPCS1-and CePCS3expressing tobacco[J]. J Plant Physiol,2010,167(12):?981-988.
[8]  ?Meyer C L,Peisker D,Courbot M,et al. Isolation and?characterization of Arabidopsis halleri and Thlaspi caerulescens phytochelatin synthases[J]. Planta,2011,234?(1):83-95.
[9]  Brunetti P,Zanella L,Proia A,et al. Cadmium tolerance?and phytochelatin content of Arabidopsis seedlings overexpressing the phytochelatin synthase gene AtPCS1[J]. J?Exp Bot,2011,62(15):5509-5519.
[10]  Zhu Y L,Pilon-Smiths E A H,Tarun A S,et al. Overexpression of glutathione synthetase in Indian mustard enhances cadmium accumulation and tolerance[J]. Plant?Physiol,1999,119:73-79.
[11]  Gisbert C,Ros R,De Haro A,et al. A plant genetically?modified that accumulates Pb is especially promising for?phytoremediation[J]. Biochem Biophys Res Commun, 2003,303:440-445.
[12]  范洪黎 . 苋菜超积累镉的生理机制研究[D]. 北京:?中国农业科学院研究生院,2007.
[13]  Zhao F J,Jiang R F,Dunham S J,et al. Cadmium uptake, translocation and tolerance in the hyperaccumulator Arabidopsis halleri[J]. New Phytol,2006,172:646-654.
[14]  ?Garbisu C,Alkorta I. Phytoextraction: a cost-effective?plant-based technology for the removal of metals from?the environment[J]. Bioresource Technoi,2001,77?(3):229-236.
[15]  Wang H C,Wu J S,Chia J C,et al. Phytochelatin synthase is regulated by protein phosphorylation at a threonine residue near its catalytic site[J]. J Agric Food?Chem,2009,57(16):7348-7355.
[16]  Vestergaard M,Matsumoto S,Nishikori S,et al. Chelation?of cadmium ions by phytochelatin synthase:role of the?cysteine-rich C-terminal[J]. Anal Sci,2008,24(2):277?-281.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133