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OALib Journal期刊
ISSN: 2333-9721
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不同方法对甘薯块根可溶性蛋白的提取效果
DOI: 10.3724/SP.J.1145.2014.00211, PP. 211-216
Keywords: 甘薯块根,蛋白质制备,sds-page,双向电泳,蛋白质组学
Abstract:
为建立一套适宜甘薯块根蛋白质组学分析的双向电泳体系（2-de），采用匀浆法、丙酮沉淀法、tca-丙酮沉淀法和酚提取法来制备甘薯块根可溶性总蛋白，并分别通过蛋白质定量、sds-page和2-de技术来评估这4种蛋白制备方法提取甘薯块根蛋白质的优劣.结果显示，每0.5g鲜薯块根，经bradford方法测定蛋白质的得率分别为1.6mg、2.4mg、1.2mg、1.9mg.sds-page的结果表明，匀浆法、丙酮法制备的蛋白质分离到的条带较少，且在约mr20×103处的储藏蛋白含量较高；tca-丙酮法和酚提取法制备的蛋白质分离到的条带则明显多于前两种方法，且mr20×103处储藏蛋白的相对含量也明显减少.2-de结果显示，匀浆法制备的蛋白质在进行双向电泳分析时，几乎不能完成聚焦过程，仅在凝胶的碱性端出现少量的蛋白质斑点，丙酮法和tca-丙酮法要优于匀浆法，但是蛋白质斑点数量仍然较少，酚提取法提取的蛋白质在进行双向电泳时分离到的蛋白质斑点数最多，为1120个，且在凝胶上均匀分布，背景清晰.综上所述，酚提取法是适宜甘薯块根蛋白质双向电泳样品制备的最佳方法.
| [1] | 1陶向,张勇为,姜玉松,王海燕,张义正.甘薯块根储藏过程中的淀粉含量变化.应用与环境生物学报,2010,16:741-744[taox,zhangyw,tongy.changesinrootstarchcontentsofsweetpotatocultivarsduringstorage.chinjapplenvironbiol,2010,16:741-
|
| [2] | 2陶向,童英,张勇为,姜玉松,王海燕,张义正.甘薯鲜薯不同储存期呼吸速率的测定与分析.四川大学学报(自然科学版),2010,47(5):1131-1136[taox,tongt,zhangyw,jiangys,wanghy,zhangyz.determinationandanalysisofsweetpotatorespiratoryrateduringstorageperiod.jsichuanuniv(natscied),2010,47(5):1131-
|
| [3] | 3张彩霞,谢高地,李士美,盖力强中国甘薯乙醇的资源潜力及空间分布.资源科学,2010,32(3):505-511[zhangcx,xiegd,lism,gailq.resourcepotentialofsweetpotatoethanolanditsspatialdistributioninchina.resoursci,2010,32:505-
|
| [4] | 4李鹏霞,王炜,胡花丽,王毓宁,刘春泉.低温贮藏下两种甘薯的品质变化研究.江西农业学报,2009,21(4):73-75[lipx,wangw,huhl,wangyn,liucq.changesofqualityoftwovarietiesofsweetpotatostoredatlowtemperature.actaagricjiangxi,2009,21(4):73-
|
| [5] | 10o’farrellph.highresolutiontwo-dimensionalelectrophoresisofproteins.jbiolchem,1975,250:4007-4021
|
| [6] | 11hollowaypj,arundelph.high-resolutiontwo-dimensionalelectrophoresisofplantproteins.analbiochem,1988,172:8-15
|
| [7] | 12mullerec,schumannm,rickersa,bommertk,wittmann-lieboldb,ottoa.studyofburkittlymphomacelllineproteinsbyhighresolutiontwo-dimensionalgelelectrophoresisandnanoelectrospraymassspectrometry.electrophoresis,1999,20:320-330
|
| [8] | 14杨艳华,崔为同,刘晓勇,朱克明,陈克平.水稻叶片蛋白质组双向电泳实验条件的改进.应用与环境生物学报,2012,18(5):819-823[yangyh,cuiwt,liuxy,zhukm,chenkp.improvementoftwo-dimensionalelectrophoresisofimprovementoftwo-dimensionalelectrophoresisofrice(oryzasatival.)proteomics.chinjapplenvironbiol,2012,18(5):819-
|
| [9] | 15bradfordmm.arapidandsensitivemethodforthequantitationofmicrogramquantitiesofproteinutilizingtheprincipleofprotein-dyebinding.analbiochem,1976,72:248-254
|
| [10] | 19刘伟霞,潘映红.适用于小麦叶片蛋白质组分析的样品制备方法.中国农业科学,2007,40(10):2169-2176[liuwa,panyh.samplepreparationmethodssuitableforwheatleafproteomeanalysis.sciagricsin,2007,40(10):2169-
|
| [11] | 21hariv.amethodforthetwo-dimensionalelectrophoresisofleafproteins.analbiochem,1981,113:332-335
|
| [12] | 22罗泽宇,杨粤军,刘选明.拟南芥蛋白质组研究中双向电泳技术条件的优化.激光生物学报,2008,17选择(4):539-545[luozy,yangyj,liuxm.optimizationontwo-dimensionalelectrophoresisforproteomicstudyinarabidopsisthaliana.actalaserbiolsin,2008,17(4):539-545
|
| [13] | 23gilmorejm,washburnmp.advancesinshotgunproteomicsandtheanalysisofmembraneproteomes.jproteomics,2010,73:2078-2091
|
| [14] | 5谢逸萍,李洪民,王欣.贮藏期甘薯块根淀粉酶活性变化趋势.江苏农业学报,2008,24(4):406-409[xieyp,lihm,wangx.changesofamylaseactivitiesinsweetpotatorootsduringstorage.jiangsujagricsci,2008,24(4):406～409
|
| [15] | 6ishigurok,yaharas,yoshimotom.changesinpolyphenoliccontentandradical-scavengingactivityofsweetpotato(ipomoeabatatasl.)duringstorageatoptimalandlowtemperatures.jagricfoodchem,2007,55:10773-10778
|
| [16] | 7luanys,zhangj,liudm,liwl.molecularcharacterizationofsweetpotatoleafcurlvirusisolatefromchina(splcv-cn)anditsphylogeneticrelationshipwithothermembersofthegeminiviridae.virusgenes,2007,35:379-385
|
| [17] | 8liangy,srivastavas,rahmanmh,strelkovse,kavnnv.proteornechangesinleavesofbrassicanapusl.asaresultofsclerotiniasclerotiorumchallenge.jagricfoodchem,2008,56:1963-1976
|
| [18] | 9aghaeik,ehsanpouraa,komatsus.proteomeanalysisofpotatoundersaltstress.jproteomeres,2008,7:4858-4868
|
| [19] | 13hajduchm,rakwalr,agrawalgk,yonekuram,pretovaa.high-resolutiontwo-dimensionalelectrophoresisseparationofproteinsfrommetal-stressedrice(oryzasatival.)leaves:drasticreductions/fragmentationofribulose-1,5-bisphosphatecarboxylase/oxygenaseandinductionofstress-relatedproteins.electrophoresis,2001,22:2824-2831
|
| [20] | 16herbertb.advancesinproteinsolubilisationfortwo-dimensionalelectrophoresis.electrophoresis,1999,20:660-663
|
| [21] | 17chenw,jijg,xuxm,hesz,rubg.proteomiccomparisonbetweenhumanyoungandoldbrainsbytwo-dimensionalgelelectrophoresisandidentificationofproteins.intjdevneurosci2003,21:209-216
|
| [22] | 18neuhoffvsr,eiblh.clearbackgroundandhighlysensitiveproteinstainingwithcoomassiebluedyesinpolyacrylamidegels.electrophoresis,1985,6:427-448
|
| [23] | 20canovasfm,dumas-gaudote,recorbetg,jorrinj,mockhp,rossignolm.plantproteomeanalysis.proteomics,2004,4:285-298
|
| [24] | 24gorga,postelw,domscheita,gunthers.two-dimensionalelectrophoresiswithimmobilizedphgradientsofleafproteinsfrombarley(hordeumvulgare):method,reproducibilityandgeneticaspects.electrophoresis,1988,9:681-692
|
| [25] | 25王小丽,gonzalezperezp,叶俊,黄丹枫.土壤宏蛋白质组学蛋白质提取方法及其应用.应用与环境生物学报,2012,18(4):691-696[wangxl,gonzalezperezpablo,yej,huangdf.methodsofproteinextractionfromsoilandtheirpotentialapplicationinstudyofsoilmetaproteome.chinjapplenvironbiol,2012,18(4):691-
|
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