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pCAMBIAvectors国际农业分子生物学应用中心-CAMBIA是一个独立的、非营利的国际研究组织,注册在澳洲的首都。主要是对农业分子生物学的研究。
1.Introduction1.1Whyanewsetofvectors?Planttransformationisnowroutineinhundredsoflaboratoriesworldwide,usingbothAgrobacteriumordirectmethods,notablyparticlebombardment.RecentlyithasbecomeclearthateventhemajorityofmonocotsaresusceptibletoefficientAgrobacterium-mediatedtransformation(forrice,seeHieietal.1997).However,thevectorsusedvarywidelybetweenlaboratoriesmakingcomparisonsofresultsbetweengroupsdifficult,andmostare‘historical’relicswithmanysubstandardfeaturesthatmakeadvancedDNAconstructionsawkwardorcumbersome.
Thesesubstandardfeaturesofmostvectorsinclude:low-copyoriginofreplicationresultinginlowyieldDNApreps;unstablereplicons,causingvariablelossoftheplasmidduringpropagation;largesize;lackofconvenientrestrictionsitesformanipulation;limitedchoiceofselectablemarkersforbothbacteriaandplants;lackofsimplewaystoconstructreportergenefusions;incompletelysequencedregions;nopathfor‘upgrades’.
Asintellectualpropertyrestrictionsbecomemoreonerous,itiscleartheabilitytoremovesegmentsofDNAwhicharenotreadilylicensed,orforwhichfreedom-to-operatecannotbeobtained,isofgreatimportanceforcommercialapplications.Therefore,developinga‘modular’structurethatencompassesanupgradepathisofthehighestpriority.Thus,thereisstillapressingneedtohaveasinglesetofvectorswhichcanbeusedforroutineandefficienttransformationofmostcrops,bothdicotandmonocot,withnumerousgeneticselections,includingroutestosimplenovelselectionupgrades.Thesevectorsshouldalsofacilitateadvancedconstruction,modificationandanalysisoftransgenesandvariants.
CAMBIAhasconstructedanewsetofvectorstomeettheserequirementsforitsownin-houseuse,andhasnowmadetheseavailabletotheresearchcommunity.Thestartingmateria
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