Description:
CreRecombinaseisaTypeItopoisomerasefrombacteriophageP1thatcatalyzesthesite-specificrecombinationofDNAbetweenloxPsites(1).TheenzymerequiresnoenergycofactorsandCre-mediatedrecombinationquicklyreachesequilibriumbetweensubstrateandreactionproducts(2).TheloxPrecognitionelementisa34basepair(bp)sequencecomprisedoftwo13bpinvertedrepeatsflankingan8bpspacerregionwhichconfersdirectionality(3).RecombinationproductsdependonthelocationandrelativeorientationoftheloxPsites.TwoDNAspeciescontainingsingleloxPsiteswillbefused.DNAbetweendirectlyrepeatedloxPsiteswillbeexcisedincircularformwhileDNAbetweenopposingloxPsiteswillbeinvertedwithrespecttoexternalsequences.

Highlights
- Isolatedfromarecombinantsource
- ExcisionofDNAbetweentwoloxPsites
- FusionofDNAmoleculescontainingloxPsites
- InversionofDNAbetweenloxPsites
ProductSource
PurifiedfromanE.colistraincarryingaplasmidencodingCreRecombinasefrombacteriophageP1withadditionalN-terminalAlaandGlyresidues(4).ReagentsSupplied
Thefollowingreagentsaresuppliedwiththisproduct:
| Storeat(°C) | Concentration | |
| CreRecombinaseReactionBuffer | -20 | 10X |
| pLox2+(linearized) | -20 | 125ng/μl |
Notes:
IncubationoftheCreRecombinasereactionmixat70°Cfor10minutesisrecommendedbeforeagarosegelanalysis.BecausetheCreRecombinasereactionisanequilibriumreaction,weobserve20-30%recombinationonourloxP2+controlsubstrate(Fig.1).Thismodestyieldproducesafaintbandonanethidiumbromidestainedgelandtheconcomitantreductioninsubstratestainingintensity.Longerincubationtimeswillnotimproverecombination,andinstead,willlikelyleadtohighermolecularweightrecombinationproducts.IncreasingtheamountofCreRecombinaseinthereactioncaninhibitrecombinationbyformingloxPdependentCre-DNAaggregates.ControlDNA:LinearizedpLox2+is3,625bpinlength,withaloxPsiteapproximately400bpfromeachend.BetweentheloxPsiteslieanoriginofreplicationandampicillin-resistancegene.RecombinationbetweentheseloxPsitesproducesacircular(2,787bp),ampicillin-resistantplasmid(whichmigratesatapproximately1.7kbona0.8%agarosegel)andone838bpDNAfragment.ebiomall.com
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基因重组只是控制同一性状表达不同而已,但是它不会改变性状的基本情况。如:控制眼睛颜色的基因,亲本的基因重组之后可能亲本为黑色,子代便为蓝色。但绝对不会控制眼睛颜色的基因在重组之后就变成控制耳朵的基因。
生物合成人胰岛素注射液,比如诺和灵R笔芯。是单一品种、不是混合的胰岛素。是不含鱼精蛋白的短效胰岛素。
提供亲和纯化相关资料:http://wenku.baidu.com/view/ef8f3831f8c75fbfc67db276
如有帮助,请采纳,谢谢~
▷ 重组蛋白生产平台
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