TheBiophysical Society of Japan General IncorporatedAssociation RNAP holoenzyme compnsing RNAP core enzyme (two a]pha gubunits one beta subunit ene beta subumt and one emega subunit) and one ot several regulators (sigma taLtors) We eonsLructed Lhe plasmid contaming three gene ot holoenzyme (two of rpoA one of rpoB dnd one of rpoC) ancl purified from E coh usmg Ni NTA column and ion exchange column Sigma subunit was a]so punfied from E co]i Combimng these purincd proteins we obtatned guficient

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... ount of RNAP In the prescntatien we will discuss the ddta of applymg pumhcd RNAP mto oul in vitTo cel] free trdnsldtion systun PURE s}stcrn 2P129 NMR eMLIte Irelp vaS6 HACI mRNA openuaMopneut NMR analysib of Irclp induced HACI mRNA processmg Ikumi Kavfal]araL ? KmcbiroHarmtil CbnyiroKolima] i Yeshivuki Tdi]akdl (lliibotcrtor} ol WoieL ulmJ lranpt)rmanon Graduare School ofPhannaceun[alScten(ei Tbhoku Ln:Ttrtttl 2Laboraton of Etcy)J ± yries eiaduate gchoot ofBioio;scal S"tn" Aaffi lnsttsurL oj Scf"lce and l"lnotos} 3hLhfnul{ Jot Prete nReserLh Osakn Un/vets/-)