中度嗜盐菌Halomonassp耐盐相关DNA片段的克隆（完整版）实用资料(可以直接使用，可编辑完整版实用资料，欢迎下载）应用与环境生物学报2006,12(3:375～378ChinJApplEnvironBiol=ISSN10062687X2006206225中度嗜盐菌Halomonassp.BYS21耐盐相关DNA片段的克隆3洪青1,2徐剑宏1王云端1武俊1张晓舟1李顺鹏133(1南京农业大学生命科学学院/农业部农业环境微生物工程重点开放实验室南京210095(2中国科学院南京土壤研究所/土壤与农业可持续发展国家重点实验室南京210008摘要通过固体亚硝基胍诱变获得了Halomonassp.BYS21的盐敏感突变株BYS21M.以22为载体在E.coliDH5α中构建了BYS21的基因文库.以文库菌E.coliDH5α(2X为辅助菌、BYS21M为受体菌进行三亲接合,pHR23,酶切确定其插入的耐盐相关DNA片段大小分为5.4耐盐相关基因的克隆奠定了基础.图5表1参14关键词中度嗜盐菌;CLCQ785:105CingofaSalt2toleranceRelatedDNAFragmentofaModerateHalophilicBacteriumHalomonassp.BYS213HONGQing1,2,XUJianhong1,WANGYunduan1,WUJun1,ZHANGXiaozhou1&LIShunpen133(1KeyLaboratoryofMicrobiologicalEngineeringAgriculturalEnvironment,MinistryofAgriculture,CollegeofLifeSciences,NanjingAgriculturalUniversity,Nanjing210095,China(2StateKeyLaboratoryofSoilandSustainableAgriculture,InstituteofSoilScience,ChineseAcademyofScience,Nanjing210008,ChinaAbstractAsalt2sensitivemutantBYS21MofstrainHalomonassp.BYS21wasobtainedbynitriguanidinemutagenesis.ThegenelibraryofBYS21wasconstructedinE.coliDH5αwithpBBR1MCS22asthevectorbythemethodofshotguncloning.Tri2parentalconjucationwasconductedwithBYS21Masrecipient,thelibrarystrainE.coliDH5α(pBBR2XasdonorsandE.coliWD803(pRK2021ashelper.Inthisway,thegenelibraryofBYS21wastransferredintoBYS21Mtolookforthecomple2mentationofthemutant.OnehybridnamedHR223,whichhadrecoveredthesamesalt2tolerancecharacteristicsastheoriginalstrainBYS21wasobtained.TherecombinantplasmidpHR223wasextractedfromtheHR223andsubjectedtorestrictionanaly2sis;theresultsshowedthatthelengthofinsertedsalt2tolerancerelatedDNAfragmentinpHR223was5.4kb.Thisfragmentlaidthefoundationforthecloningofsalt2relatedgeneofHalomonassp.BYS21.Fig5,Tab1,Ref14Keywordsmoderatelyhalophilicbacteria;salt2tolerancerelatedDNAfragment;cloningCLCQ785:Q939.105盐单胞菌(Halomonas是一类耐盐性很强的细菌,可在0.5%～32.5%的NaCl条件下生长,属于中度嗜盐菌(moderatehalophiles.它们和极端嗜盐古菌(extremehalophiles都属于极端微生物[1],但比嗜盐古菌分布更广泛,对营养要求低,更易于适应环境[2～5].盐单胞菌独特的生物学特性、生理机制和遗传机制,特别是渗透