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1、第15卷第5期:47生物技术Vol115,No15:472005年10月BIOTECHNOLOGYOct12005立了一种PCR方法,通过扩增致病性蜡样芽孢杆菌的hblA基因来实现对蜡样芽胞杆菌的快速检测。实验结果表明,以hb2lA基因为目的基因设计引物建立的PCR检测方法能特异扩增蜡样芽孢杆菌的hblA基因,且与其他的菌株均交叉反应,其检测敏感性可达9CFUPml,该方法具有良好的应用前景,其快速、灵敏度高、特异性强,省时省力,易于在实际检验工作中推广应用。参考文献:[1]VesaMantynen,KristinaLindstrom.ArapidPCR
2、-basedDNAtestforen2terotoxicBacilluscereus[J].AppliedandEnvironmentalMicrobiology,1998,64(5):1634-1639.[2]IroshiFukushima,YoshieTsunomori,RyotaroSeki.Duplexreal-timeSYBRgreenPCRassaysfordetectionof17speciesoffoodorwaterbornepatho2图2不同稀释浓度的蜡样芽孢杆菌经hblA引物扩增结果的电泳图gensinstools[J].Jour
3、nalofClinicalMicrobiology,2003,41(11):5134-(从左到右的菌数分别为9.2×10543210)、10、10、10、10、105146.Fig.2AmplificationfragmentsofdifferentdilutionconcentrationB.cereus[3]HJBach,DErrampalli,KTLeung,etal.Specificdetectionofthegenefor(fromlefttorighttheconcentrationofB.cereusis9.2×105432,10,10,1
4、0,theextracellularneutralproteaseofBacilluscereusbyPCRandblothybridiza21010,10CFUPmlrespectively)tion[J].AppliedandEnvironmentalMicrobiology,1999,65(7):3226-3228.[4]BjarneMunkHansen,NielsBohseHendriksen.DetectionofenterotoxicBa2cilluscereusandBacillusthuringiensisstrainsbyPCRanal
5、ysis[J].AppliedandEnvironmentalMicrobiology,2001,67(1):185-189.[5]ShoichiYamada,EijiOhashi,NorioAgata,etal.CloningandnucleotidesequenceanalysisofgyrBofBacilluscereus,B.thuringiensis,B.mycoides,andB.anthracisandtheirapplicationtothedetectionofB.cereusinrice[J].Ap2pliedandEnvironme
6、ntalMicrobiology,1999,65(4):1483-1490.[6]Yu-HsiuChang,Yung-HuiShangkuan,Hung-ChiLin,etal.PCRas2sayofthegroELgenefordetectionanddifferentiationofBacilluscereusgroupcells[J].AppliedandEnvironmentalMicrobiology,2003,69(8):4502-4510.图3hblA基因用于实际样品的检测的扩增片段[7]MonikaEhling-Schulz,Martin
7、aFricker,SiegfriedScherer.Identification(图中1、5、6、8为致病性腊样芽孢杆菌阳性样品其他为阴性样品)ofemetictoxinproducingBacilluscereusstrainsbyanovelmolecularassay[J].Fig.3AmplificationfragmentsofdifferentsamplesFEMSMicrobiologyLetters,2004,23(2):189-195.3讨论[8]ElseMarieFykse,JaranStrandOlsen,GunnarSkogan.
8、Applicationofsoni2由于蜡样芽孢杆菌及其芽孢广泛存在于周围的环境