分子生物学实验指导

12−SouthernMAASBDSBDWesternSBD34__482.5≥≥567°°89α10µ°α°°µµµµµµµ°µ°µµ°11β12°°°µµµµµµ°13µµµµ°µ1415×××µ×°165148497317′′°°αβ′′αββαβααβββββαβαβ18αλ°µ°µµµµµµ°λµµµ19µ°µ°°µµµ°µµµµµµµ°µ°µµµ°°20°21λλ22°µ°°λ°µ°°°°µµµ23λ24ββ25µµ26°27××°°°°µµ28°µ°28srRNA18srRNAµµµµµ29⎯′′′′′′′′′′′µµ30µµµµ31µµ°°°°µµµµµµµµµµµµµµµµµ32°µ°°33µµµµ°°34′′35µµµµµµ°°°°°µ36µµ37ββ38°°µµµ°µ°−39°°°°β40µµµµµµµ°µ4142−4344µ°4546SouthernSouthernSouthernSouthernnitrocellulosefiltermembraneNCSouthernSouthernSouthernSouthern°α47µµµ48µ49°µµµµµαµµµµµµµFicoll50Southernµµµµµµµµµµ51SouthernSouthern52MAAMAA-DNA1.(2004):PCR,pp90-92,.53SBD:18pTrcHisB(Bacilluscirculansstrain251)(cyclodextringlycosyltransferase,CGTase)(starch-bindingdomain,SBD)E.coliTop10DNAT4DNA(CGTase)(SBD)pTrcHisBTop10DNApTrcHisB/SBDDNA1.(2004),.2.Lawson,C.L.,vanMontfort,R.,Strokopytow,B.,Rozeboom,H.J.,Kalk,K.H.,deVries,G.E.,Penninga,D.,Dijkhuizen,L.andDijkstra,B.W.(1994)NucleotidesequenceandX-raystructureofcyclodextringlycosyltransferasefromBacilluscirculansstrain251inamaltose-dependentcrystalform.J.Mol.Biol.236,590-600.3.Ji,Q.,lVincken,J.-P.,Suurs,L.C.J.M.and.Visser,R.G.F.(2003)Microbialstarch-bindingdomainsasatoolfortargetingproteinstogranulesduringstarchbiosynthesis.PlantMol.Biol.51,789-801.4.(PhastSystem)(ProBondResin)(RainBow),NAP-25()12.5%SDS-PAGEIPTG(-β-D-)SBDSDS-PAGESBD1.(2004):,pp93-97,.2.QinJi,Jean-PaulVincken,LucC.J.M.Suurs,andRichardG.F.Visser(2003)Microbialstarch-bindingdomainsasatoolfortargetingproteinstogranulesduringstarchbiosynthesis.PlantMol.Biol.51,789-801.3.(PhastSystem),12.5%SDS-PAGE(50mm×50mm×3mm),()SBDHybondECLSBD()IgG()SBDSDS1.(2004):,pp98-101,.4.QinJi,Jean-PaulVincken,LucC.J.M.Suurs,andRichardG.F.Visser(2003)Microbialstarch-bindingdomainsasatoolfortargetingproteinstogranulesduringstarchbiosynthesis.PlantMol.Biol.51,789-801.566Western(PhastSystem),12.5%SDS-PAGE(50mm×50mm×3mm),()SBDSBD(Trypsin)HybondECLSBD()IgG()SBD,SDS1.(2004):,pp98-101,.5.QinJi,Jean-PaulVincken,LucC.J.M.Suurs,andRichardG.F.Visser(2003)Microbialstarch-bindingdomainsasatoolfortargetingproteinstogranulesduringstarchbiosynthesis.PlantMol.Biol.51,789-801.5781.J.[]()()22001DNA.3[]()()22001DNA.3ŵµµµµµµµµµµµµµ6061°°°°°°62°°×°××·63××°°°°ºµµºµµºµµºµµºµµºµµµ64°6566×××××××××××67×°6869µ70°°°°