HUANG Shan,WANG Wei-qi,HOU Wen-sheng.Cloning and Expression Analysis of Usp1 Gene from Soybean[J].Soybean Science,2012,31(04):546-551.[doi:10.3969/j.issn.1000-9841.2012.04.008]
大豆Usp1基因的克隆和表达分析
- Title:
- Cloning and Expression Analysis of Usp1 Gene from Soybean
- 文章编号:
- 1000-9841(2012)04-0546-06
- Keywords:
- Soybean; UspA; Sequence analysis; Gene expression
- 分类号:
- S565.1
- 文献标志码:
- A
- 摘要:
- 在对大豆根系盐胁迫抑制差减杂交文库EST序列分析的基础上,利用RT-PCR技术克隆得到了一个大豆UspA基因,命名为GmUsp1,其编码一个包含164个氨基酸残基的多肽链。多序列比对和编码蛋白结构分析表明,GmUsp1属于泛应激蛋白(Universal Stress Protein)家族成员之一。其氨基酸序列与蚕豆Vf_enod18序列相似度最高,属于MJ-0577类中含有ATP结合位点的UspA亚族,与MJ-0577有相似的蛋白二级结构。分别采用250 mmol·L-1 NaCl、100 μmol·L-1 ABA和30% PEG 6000进行胁迫处理,耐盐品种文丰7号和盐敏感品种Union的GmUsp1均被诱导表达,二者对胁迫诱导响应时间和表达量存在差异,说明GmUsp1可能参与大豆对非生物逆境胁迫的应答调控。
- Abstract:
- According to the EST sequences of the Suppression Subtractive Hybridization(SSH)cDNA libraries,a soybean UspA gene named by GmUspl was cloned from the soybean by RT-PCR.The complete cDNA of GmUsp1 was 1 047 bp,it encoded a peptide which contained 164 amino acid residues.The 495 bp CDS showed that it belonged to the universal stress protein(UspA)family,by using the BLASTp tool of NCBI database.Homology analysis showed GmUsp1 had the highest similarity with the bean Vf_enod18,and it belonged to the group of UspA,which represented by the ATP-binging structure of the MJ-0577 protein from Methanococcus jannaschii.It also had the similar protein secondary structure with MJ-0577.In the salt sensitive variety Union and tolerant variety Wenfeng No.7,the expression of GmUsp1 was induced by 250 mmol·L-1 NaCl,100 μmol·L-1 ABA and 30% PEG-600 treatment,however,it had different response time and expression level.So GmUsp1 might be involved in plant stress adaptation,it suggested that GmUsp1 may play a role in salt stress adaptation in soybean.
参考文献/References:
[1]Nystrom T,Neidhardt F C.Expression and role of the universal stress protein,UspA,of
Escherichia coli
?during growth arrest[J].Molecular Microbiology,1994,11:537-544.
[2]Nystrom T,Neidhardt F C.Isolation and properties of a mutant of
Escherichia coli
?with an insertional inactivation of the
UspA
?gene,which encodes a universal stress protein[J].Journal of Bacteriology,1993,175:3949-3956.
[3]Liu W T,Karavolos M H,Bulmer D M,et al.Role of the universal stress protein UspA of Salmonella in growth arrest,stress and virulence[J].Microbial Pathogenesis,2007,42:2-10.
[4]Freestone P,Nystrom T,Trinei M,et al.The universal stress protein,UspA,of
Escherichia coli
?is phosphorylated in response to stasis[J].Journal of Molecular Biology,1997,274:318-324.
[5]Zarembinski T I,Hung L W,Mueller-Dieckmann H J,et al.Structure-based assignment of the biochemical function of a hypothetical protein:a test case of structural genomics[J].Proceedings of the National Academy of Sciences of the United States of America,1998,95:15189-15193.
[6]Sousa M C,McKay D B.Structure of the universal stress protein from
Haemophilus influenzae
[J].Structure,2001,9:1135-1141.
[7]David K,Joshua B,Douglas W S,et al.
Arabidopsis
?proteins containing similarity to the universal stress protein domain of bacteria[J].Plant Physiology,2003,131:1209-1219.
[8]Wang W Q,Li L,Huang S,et al.A secondary suppression subtractive hybridization method for isolation and identification of some salt-induced genes in soybean(
Glycine max
?L.Merr.)[J].Australian Journal of Crop Science,2012,6:46-55.
[9]李亮,侯文胜.抑制差减杂交技术在大豆研究中的应用[J].大豆科学,2010,29(4):702-706.(Li L,Hou W S.Application of suppression subtractive hybridization in soybean research[J].Soybean Science,2010,29(4):702-706.)
[10]常汝镇,陈一舞,邵桂花,等.盐对大豆农艺性状及籽粒品质的影响[J].大豆科学,1994,13(2):101-105.(Chang R Z,Chen Y W,Shao G H,et al.Effect of salt on agricultural characters and chemical quality of seed in soybeans[J].Soybean Science,1994,13(2):101-105.)
[11]郭蓓,邱丽娟,邵桂花,等.大豆耐盐基因的PCR标记[J].中国农业科学,2000,33(1):10-16.(Guo B,Qiu L J,Shao G H,et al.Tagging salt tolerant gene using PCR markers in soybean[J].Scientia Agricultura Sinica,2000,33(1):10-16)
[12]Jian B,Liu B,Bi Y,et al.Validation of internal control for gene expression study in soybean by quantitative real-time PCR[J].BioMed Central:Molecular Biology,2008,9:59.
[13]Kenneth J L,Thomas D S.Analysis of relative gene expression data using Real-Time Quantitative PCR and the 2-ΔΔCT method[J].Methods,2001,25:402-408.
[14]Hohnjec N,Kuster H,Albus U,et al.The broad bean nodulin VfENOD18 is a member of a novel family of plant proteins with homologies to the bacterial MJ0577 superfamily[J].Molecular and General Genetics,2000,264:241-250.
[15]Chou M X,Wei X Y,Chen D S,et al.A novel nodule-enhanced gene encoding a putative universal stress protein from
Astragalus sinicus
[J].Journal of Plant Physiology,2007,164 :764-772.
[16]Maqbool A,Zahur M,Husnain T,et al.GUSP1 and?GUSP2,two drought-responsive genes in?Gossypium arboreum?have homology to universal stress proteins[J].Plant Molecular Biology Reporter,2009,27:109-114.
[17]Sauter M,Rzewuski G,Marwedel T,et al.The novel ethylene-regulated gene?
OsUsp
1 from rice encodes a member of a plant protein family related to prokaryotic universal stress proteins[J].Journal of Experimental Botany ,2002,53:2325-2331.
[18]Nystrom T,Neidhardt F C.
Effects of overproducing the universal stress protein UspA,in Escherichia coli K-12[J].Journal of Bacteriology,1996,178:927-930.
[19]Zegzouti H,Jones B,Frasse P,et al.Ethylene-regulated gene expression in tomato fruit:characterization of novel ethylene-responsive and ripening-related genes isolated by differential display[J].Plant Journal,1999,18:589-600.
[20]Zhu J K.Plant salt tolerance[J].Trends in Plant Science,2001,6(2):66-71.
相似文献/References:
[1]刘章雄,李卫东,孙石,等.1983~2010年北京大豆育成品种的亲本地理来源及其遗传贡献[J].大豆科学,2013,32(01):1.[doi:10.3969/j.issn.1000-9841.2013.01.002]
LIU Zhang-xiong,LI Wei-dong,SUN Shi,et al.Geographical Sources of Germplasm and Their Nuclear Contribution to Soybean Cultivars Released during 1983 to 2010 in Beijing[J].Soybean Science,2013,32(04):1.[doi:10.3969/j.issn.1000-9841.2013.01.002]
[2]李彩云,余永亮,杨红旗,等.大豆脂质转运蛋白基因GmLTP3的特征分析[J].大豆科学,2013,32(01):8.[doi:10.3969/j.issn.1000-9841.2013.01.003]
LI Cai-yun,YU Yong-liang,YANG Hong-qi,et al.Characteristics of a Lipid-transfer Protein Gene GmLTP3 in Glycine max[J].Soybean Science,2013,32(04):8.[doi:10.3969/j.issn.1000-9841.2013.01.003]
[3]王明霞,崔晓霞,薛晨晨,等.大豆耐盐基因GmHAL3a的克隆及RNAi载体的构建[J].大豆科学,2013,32(01):12.[doi:10.3969/j.issn.1000-9841.2013.01.004]
WANG Ming-xia,CUI Xiao-xia,XUE Chen-chen,et al.Cloning of Halotolerance 3 Gene and Construction of Its RNAi Vector in Soybean (Glycine max)[J].Soybean Science,2013,32(04):12.[doi:10.3969/j.issn.1000-9841.2013.01.004]
[4]张春宝,李玉秋,彭宝,等.线粒体ISSR与SCAR标记鉴定大豆细胞质雄性不育系与保持系[J].大豆科学,2013,32(01):19.[doi:10.3969/j.issn.1000-9841.2013.01.005]
ZHANG Chun-bao,LI Yu-qiu,PENG Bao,et al.Identification of Soybean Cytoplasmic Male Sterile Line and Maintainer Line with Mitochondrial ISSR and SCAR Markers[J].Soybean Science,2013,32(04):19.[doi:10.3969/j.issn.1000-9841.2013.01.005]
[5]卢清瑶,赵琳,李冬梅,等.RAV基因对拟南芥和大豆不定芽再生的影响[J].大豆科学,2013,32(01):23.[doi:10.3969/j.issn.1000-9841.2013.01.006]
LU Qing-yao,ZHAO Lin,LI Dong-mei,et al.Effects of RAV gene on Shoot Regeneration of Arabidopsis and Soybean[J].Soybean Science,2013,32(04):23.[doi:10.3969/j.issn.1000-9841.2013.01.006]
[6]杜景红,刘丽君.大豆fad3c基因沉默载体的构建[J].大豆科学,2013,32(01):28.[doi:10.3969/j.issn.1000-9841.2013.01.007]
DU Jing-hong,LIU Li-jun.Construction of fad3c Gene Silencing Vector in Soybean[J].Soybean Science,2013,32(04):28.[doi:10.3969/j.issn.1000-9841.2013.01.007]
[7]张力伟,樊颖伦,牛腾飞,等.大豆“冀黄13”突变体筛选及突变体库的建立[J].大豆科学,2013,32(01):33.[doi:10.3969/j.issn.1000-9841.2013.01.008]
ZHANG Li-wei,FAN Ying-lun,NIU Teng-fei?,et al.Screening of Mutants and Construction of Mutant Population for Soybean Cultivar "Jihuang13”[J].Soybean Science,2013,32(04):33.[doi:10.3969/j.issn.1000-9841.2013.01.008]
[8]盖江南,张彬彬,吴瑶,等.大豆不定胚悬浮培养基因型筛选及基因枪遗传转化的研究[J].大豆科学,2013,32(01):38.[doi:10.3969/j.issn.1000-9841.2013.01.009]
GAI Jiang-nan,ZHANG Bin-bin,WU Yao,et al.Screening of Soybean Genotypes Suitable for Suspension Culture with Adventitious Embryos and Genetic Transformation by Particle Bombardment[J].Soybean Science,2013,32(04):38.[doi:10.3969/j.issn.1000-9841.2013.01.009]
[9]王鹏飞,刘丽君,唐晓飞,等.适于体细胞胚发生的大豆基因型筛选[J].大豆科学,2013,32(01):43.[doi:10.3969/j.issn.1000-9841.2013.01.010]
WANG Peng-fei,LIU Li-jun,TANG Xiao-fei,et al.Screening of Soybean Genotypes Suitable for Somatic Embryogenesis[J].Soybean Science,2013,32(04):43.[doi:10.3969/j.issn.1000-9841.2013.01.010]
[10]刘德兴,年海,杨存义,等.耐酸铝大豆品种资源的筛选与鉴定[J].大豆科学,2013,32(01):46.[doi:10.3969/j.issn.1000-9841.2013.01.011]
LIU De-xing,NIAN Hai,YANG Cun-yi,et al.Screening and Identifying Soybean Germplasm Tolerant to Acid Aluminum[J].Soybean Science,2013,32(04):46.[doi:10.3969/j.issn.1000-9841.2013.01.011]
备注/Memo
基金项目:国家自然科学基金(30771358)。