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OALib Journal期刊
ISSN: 2333-9721
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杂合抗菌肽lpcb在毕赤酵母中的表达条件优化
DOI: 10.3724/SP.J.1145.2012.00065, PP. 65-69
Keywords: 杂合抗菌肽,lfcin-pro-cecropinb,毕赤酵母,表达条件优化
Abstract:
为获取能表达具有天然活性的杂合抗菌肽的重组酵母菌，确定其最佳表达条件，将构建好的重组酵母表达载体ppiczα-a-cecropinb（1~10）-magainin2（1~12）（简称ppiczα-a-cbma）和ppiczα-a-lfcin-pro-cecropinb（简称ppiczα-a-lpcb）经sacⅰ线性化处理，分别电转入巴斯德毕赤酵母pichiapastorisx-33，利用抗生素zeocin筛选阳性克隆，pcr扩增验证，甲醇诱导表达.采用抑菌圈法初步测定产物活性，优化杂合肽lpcb诱导时间、诱导剂浓度、诱导温度、培养基ph等表达条件，传代检测其质粒稳定性.结果显示，基因工程菌p.pastorisx-33/ppiczα-a-cbma和p.pastorisx-33/ppiczα-a-lpcb成功构建.表达产物前者抑菌活性微弱，后者抑菌活性良好.杂合肽lpcb的最佳表达条件为：25℃，ph中性培养，每24h添加0.5%（v/v，φ）甲醇，诱导表达72h.重组酵母菌ppiczα-a-lpcb在培养超过100代后，未发生质粒丢失.图4参20
| [1] | 4thomasfl,ahmadz.inhibitionofescherichiacoliatpsynthasebyamphibianantimicrobialpeptides.internjbiolmacromol,2010,46(3):367~374
|
| [2] | 5maxb(马�玻),zhangym(张玉明),nizh(倪志华),zhouyf(周艳芬).researchonthestructureandfunctionofantimicrobialpeptides.janhuiagricsci(安徽农业科学),2009,37(23):10878~10880
|
| [3] | 6lópez-garcíab,beatrizma,sansb,delosrv,manningmj,joségg,álvaromp.productionofthebiotechnologicallyrelevantafpfromaspergillusgiganteusintheyeastpichiapastoris.protexpress&purif,2010,70(2):206~210
|
| [4] | 7santosis,carvalhoao,souza-filhoga,nascimentovv,machadoolt,gomesvm.purificationofadefensinisolatedfromvignaunguiculataseeds,itsfunctionalexpressioninescherichiacoli,andassessmentofitsinsectα-amylaseinhibitoryactivity.protexpress&puri,2010,71(1):8~15
|
| [5] | 8yangs,aihx,songr,liangzn,lijf,zhangsq.expressionandpurificationofmoricincm4andhumanβ-defensins4inescherichiacoliusinganewtechnology.origresartmicrobiolres,2010,165(8):713~718
|
| [6] | 9ramosr,dominguesl,gamam.escherichiacoliexpressionandpurificationofll37fusedtoafamilyiiicarbohydrate-bindingmodulefromclostridiumthermocellum.protexpress&purif,2010,71(1):1~7
|
| [7] | 10lic,blenckehm,smithlc,karptm,stensvågk.tworecombinantpeptides,spstrongylocins1and2,fromstrongylocentrotuspurpuratus,showantimicrobialactivityagainstgram-positiveandgram-negativebacteria.dev&compimmunol,2010,34(3):86~292
|
| [8] | 14jiangx(姜昕),mag(马根),xuzc(许正超),wup(吴鹏),qiaodr(乔代蓉),xuh(徐辉),caoy(曹毅).secretedexpressionandactivitydetectionoftwohybridantimicrobialpeptidesinpichiapastoris.jsichuanunivnatsci(四川大学学报自然科学版),2007,44(3):637~678
|
| [9] | 15wangp,bangjk,kimhj,kimjk,kimy,songys.antimicrobialspecificityandmechanismofactionofdisulfide-removedlinearanalogsoftheplant-derivedcys-richantimicrobialpeptideib-amp1.peptides,2009,30(12):2144~2149
|
| [10] | 19hocquelleta,odaertb,cabannec,noubhania,dieryckw,jouclag,senechalcl,milenkovm,chaignepains,schmitterjm,claverols,santarellix,dufourcej,bonneum,garbayb,costagliolip.structure�activityrelationshipofhumanliver-expressedantimicrobialpeptide2.peptides,2010,31(1):58~66
|
| [11] | 1bolintineanud,hazratie,davisht,lehrerri,kaznessisyn.antimicrobialmechanismofpore-formingprotegrinpeptides:100porestokille.coli.peptides,2010,31(1):1~8
|
| [12] | 2mihajlovicm,lazaridist.antimicrobialpeptidesbindmorestronglytomembranepores.biochimetbiophysacta-biomembranes,2010,1798,(8):1494~1502
|
| [13] | 3wugq,wuhb,lilx,fanxb,dingjx,lixf,xit,shenzl.membraneaggregationandperturbationinducedbyantimicrobialpeptideofs-thanatin.biochemi&biophysrescommun,2010,395(1):31~35
|
| [14] | 11invitrogen.instructionmanualformulti-copypichiaexpressionkit-versionf[z].america:invitrogen,1999
|
| [15] | 12lixl(李秀兰),daizy(戴祝英),zhangsq(张双全).studyoncorrelationandcomparationbetweenthetwomethodsofdetectingbiologicalactivityofantibacterialpeptide-agarosediffusionassayandliquidphaseassay.jnanjinnormunivnatsci(南京师范大学学报自然科学版),1998,21(2):81~83
|
| [16] | 13[美]f.奥斯伯等.精编分子生物学实验指南.北京:科学出版社,1997
|
| [17] | 16fangc(方超),zhangxf(张晓凡),zhouy(周颖),baih(白卉),houz(侯征).progressinthedevelopmentandapplicationofantimicrobialpeptides.chinjantibiot(中国抗生素杂志),2010,35(11):809~813
|
| [18] | 17hej,andersonmh,shiwy,eckertr.designandactivityofa‘dual-targeted’antimicrobialpeptide.internjantimicrobagents,2009,33(6):532~537
|
| [19] | 18nanyh,bangjk,shinsy.designofnovelindolicidin-derivedantimicrobialpeptideswithenhancedcellspecificityandpotentanti-inflammatoryactivity.peptides,2009,30(5):832~838
|
| [20] | 20takahashid,shuklask,prakasho,zhanggl.structuraldeterminantsofhostdefensepeptidesforantimicrobialactivityandtargetcellselectivity.biochimie,2010,92(9):1236~1241
|
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