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OSRg OSRgoN(SignalP http://www.cbs.dtu.dk/services/signalP ) bKmǏ z-N[+T[temRNA 5 zvContigы:Nˆ}v^R; 6qT(uSignalPoN[MR50*N(lWx^R(N,{N*NATG[^v2ukx(lxMet_Y)ۏLċ0O YgSignalPRg~Qckb~g RKmՋ^R gS:NOS; GPY(WKmՋ^Rv,{N*NMet 5 zX[(W~bk[xP[ 勏^R:NOSvS'`f'Y0ZZ' >#  ' 3# # # # # ' f# ^     =- DB ~bk[xP[ ~bk[xP[: TAA, TAG,TGA GC% = 50% ~bk[xP[k 64 bpQsN!k GC% > 50% ~bk[xP[k100 200 bp QsN!k 1uNYpeWV ORF GWYN50*N[xP[ VdkgSv b^/f ORF N\N100 *N[xP[0 0PP!f   >0 C 3 zvnx 3 zvnx;N9hncPoly(A)>\^R,KmՋContig N+TPoly(A)^R R9hncR>\OS^R AATAAA TBLAST Tn'`k~gqQ T$Re0 b $f% 3% 38% 3% 3% 33   ?, zD ^x^R0Q+TP[ ؚI{w8huirYpeY>fP[^ N\N100 *N[xP[ gv N0R50*N[xP[uf\  > f1>"  @1 E [xP[OP1r'` x TN(lWxv N T[xP[y:N TIN[x vQ]+RN(W[xP[v,{3MOxW N T0 N Ty^\O(u TIN[xvs g_'Y]_ YN{|WV-N N(lxAle [xP[Y:NGCA,GCCbGCT, GCG_\O(u0$} fsh-.A2 NF Y>fP[ Q+TP[Lu Y>fP[TQ+TP[vLu gNNf>fvyr_ Y Q+TP[v5 zbyOSOMOdonor site 8^vz^:N 5 AG!GTTAAGT-3  3 zSySSOMOacceptor site), Y:N5 PyPyPyPyPyPyCAG-3 ( Py 'VvU8hx TbC)BP $fF#T1  B3 G N8nc6Rz^ QNN@b gWVbd~P[ N8n gc^R [NSNDNA~Tˆ}v\O(u c6RWVh0 SY*N+RuirvWV~yr g~b_NS\O:N$R+ROnc Y iRirWV~YWVv N8n gCpG\0 &zZtf8)@ C7 H oNKm Ǒ(uNCBIvORFKmoN ( ORF finder: http://www.ncbi.nlm.nih.gov/gorf/orfig.cgi )$ReORFvSV0HZfZ% 3 D6 B1.2 mRNAv5 zsSlU_wYMOp:S Ǐ Tn'`kegKmmRNAv5 z g8^(uvNlU_wYMOpvsQvpenc^/fw8h/TRP[penc^(The TRADAT Project , Eukaryotic Promoter Database, EPD. http://www.epd.unil.ch/ )0\ % % %% % q% DE4 1.3 Tng_ Ǐ]X[eQpenc^-NvWVz^N_gvWV~^RۏLk N-Ng~bSNKN9SMvxWz^SvQkO (uNLu[WVvely:N Tng0  ] fQYF5  Tn gY NQy`Q A DNA^RgNGrk[hQv T B _>exFhORF cR{|fP[ C _>exFhыb(lWx^RvvfP[ N T ONu}YQag^ NNvBgN&^ YgWV/fgNWV[evbXT_NOQsY*NOo` b Q~~NN'`TS6kv 4Z $ffr EH9 C WVhNir0N^v YgΘ^NO (ubNorthern BgNTRirBgNZoo-blotting Rg0 bNorthern BgN  9hnc]wvDNAz^_ir NmRNASO-NibXWVNir QNDNA:NcNKNBgN0Z     I: RirVBgN  9hncNsQ|vO?K<3.nx[DNAz^-NWVvMOn(f$ A Ǐ[hQcDNA^RvKm^0[k NSNWV~DNAvk nx[WV@b(Wv:SW B Ǐiry]^zW OVTirtVegnx[WVvMOnFFRU ld0(  r  S 0^*(dp * P  C (A 72062-1`   0`* :  8NyYe^: YO1r=N u}Tyf[b RP[uirf[NuirOo`f[| 2H  0޽h ? 33___PPT10i.T0*$+D=' = @B +  6.(  r  S *$0F  * x  c $*$]P h} *   TA $it@: p *H  0޽h ? fff80___PPT10.]t6$  t$(  tr t S D\N$0F  N r t S aN$0 N H t 0޽h ? fff80___PPT10.pr + B< mB ,? 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OSRg OSRgoN(SignalP http://www.cbs.dtu.dk/services/signalP ) bKmǏ z-N[+T[temRNA 5 zvContigы:Nˆ}v^R; 6qT(uSignalPoN[MR50*N(lWx^R(N,{N*NATG[^v2ukx(lxMet_Y)ۏLċ0O YgSignalPRg~Qckb~g RKmՋ^R gS:NOS; GPY(WKmՋ^Rv,{N*NMet 5 zX[(W~bk[xP[ 勏^R:NOSvS'`f'Y0ZZ' >#  ' 3# # # # # ' f# ^     =- DB ~bk[xP[ ~bk[xP[: TAA, TAG,TGA GC% = 50% ~bk[xP[k 64 bpQsN!k GC% > 50% ~bk[xP[k100 200 bp QsN!k 1uNYpeWV ORF GWYN50*N[xP[ VdkgSv b^/f ORF N\N100 *N[xP[0 0PP!f   >0 C 3 zvnx 3 zvnx;N9hncPoly(A)>\^R,KmՋContig N+TPoly(A)^R R9hncR>\OS^R AATAAA TBLAST Tn'`k~gqQ T$Re0 b $f% 3% 38% 3% 3% 33   ?, zD ^x^R0Q+TP[ ؚI{w8huirYpeY>fP[^ N\N100 *N[xP[ gv N0R50*N[xP[uf\  > f1>"  @1 E [xP[OP1r'` x TN(lWxv N T[xP[y:N TIN[x vQ]+RN(W[xP[v,{3MOxW N T0 N Ty^\O(u TIN[xvs g_'Y]_ YN{|WV-N N(lxAle [xP[Y:NGCA,GCCbGCT, GCG_\O(u0$} fsh-.A2 NF Y>fP[ Q+TP[Lu Y>fP[TQ+TP[vLu gNNf>fvyr_ Y Q+TP[v5 zbyOSOMOdonor site 8^vz^:N 5 AG!GTTAAGT-3  3 zSySSOMOacceptor site), Y:N5 PyPyPyPyPyPyCAG-3 ( Py 'VvU8hx TbC)BP $fF#T1  B3 G N8nc6Rz^ QNN@b gWVbd~P[ N8n gc^R [NSNDNA~Tˆ}v\O(u c6RWVh0 SY*N+RuirvWV~yr g~b_NS\O:N$R+ROnc Y iRirWV~YWVv N8n gCpG\0 &zZtf8)@ C7 H oNKm Ǒ(uNCBIvORFKmoN ( ORF finder: http://www.ncbi.nlm.nih.gov/gorf/orfig.cgi )$ReORFvSV0HZfZ% 3 D6 B1.2 mRNAv5 zsSlU_wYMOp:S Ǐ Tn'`kegKmmRNAv5 z g8^(uvNlU_wYMOpvsQvpenc^/fw8h/TRP[penc^(The TRADAT Project , Eukaryotic Promoter Database, EPD. http://www.epd.unil.ch/ )0\ % % %% % q% DE4 1.3 Tng_ Ǐ]X[eQpenc^-NvWVz^N_gvWV~^RۏLk N-Ng~bSNKN9SMvxWz^SvQkO (uNLu[WVvely:N Tng0  ] fQYF5  Tn gY NQy`Q A DNA^RgNGrk[hQv T B _>exFhORF cR{|fP[ C _>exFhыb(lWx^RvvfP[ N T ONu}YQag^ NNvBgN&^ YgWV/fgNWV[evbXT_NOQsY*NOo` b Q~~NN'`TS6kv 4Z $ffr EH9 C WVhNir0N^v YgΘ^NO (ubNorthern BgNTRirBgNZoo-blotting Rg0 bNorthern BgN  9hnc]wvDNAz^_ir NmRNASO-NibXWVNir QNDNA:NcNKNBgN0Z     I: RirVBgN  9hncNsQ|vO?K<3.nx[DNAz^-NWVvMOn(f$ A Ǐ[hQcDNA^RvKm^0[k NSNWV~DNAvk nx[WV@b(Wv:SW B Ǐiry]^zW OVTirtVegnx[WVvMOnFFRM  d\0X(  X X s *8S $"6 ?!  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Zb$mܚ @؇D lY Zb$=뛞.'Rִ>m.e Z 0e0e     A@  A5% 8c8c     ?1 d0u0@Ty2 NP'p<'pA)BCD|E||s " 0e@        @ABC DEEFGHIJK5%LMNOPQRSTUWYZ[ \]^_ `abN E5%  N E5%  N F   5%    !"?N@ABC DEFFGHIJK5%LMNOPQRSTUWYZ[ \]^_ `ab@f3@f8qʚ;<7ʚ;g4EdEdv 0pppp@ <4dddd` 0, <4!d!d 0,2l___PPT10LD[SOang0z[ 0DArial0z[ 0" DBatang0z[ 0 ppH___PPT9BpIE&#%?  %:{,{N WV~Km^N^R~ňf  + v;NQ[: NHN/fWV~ NHN/fWV DNAKm^vel DNA^Rv~ň N{|WV~R 4l;zWV~R TWV~f[26 f6f@ ,1. NHN/fWV~  A WV~1\/fN*Niry-N@b gWVvteSO~b0 WV~ g$NB\aINW Oir(TW OOo`0 c_u}TveYy 1\NteSO4ls^xvzWVvX[(W0WVv~gNR0WVKNvvNsQ|0 h " d%ff3 iUFVGWHXI ͑ Yz^ Gؚ^͑ Yz^ ^Q*N  QCS*Nbp bpeQ~v*N  N~vN*N >\vޏ 2NTcR Ɩ-NR^NgrSOvyr[:SkYz| @wN|I{ _NykSfDNA -N^͑ Yz^ N,RceNte*NWV~-N ^Tbpe]+R_'Y USNz^ WV;NMONUSNz^ Rir-NUSNz^~`S50 iir-NUSNz^~`S200dZZ0" dZ0dZ 0" dZ0dZ10" dZG8@^YJZK[L\M V3. DNAKm^vel ~bklKm^ Sf[MlKm^ ꁨRSKm^ ^8^ĉDNAKm^0 ff6   3.1 ~bklKm^(the chain termination method) W,gSt: ǏTbNUSDNANevY8hx 1uNTbvNeS(W N TMOn:g~bkS^ NuS]N*N8hxvDNARP[ N egS_KmDNARP[vz^0< f"fS# ( b/g~NBl$f $6RYUS!jg ! \US!jgNN\k_irkp ! ReQDNAYZv 4y1'l8hx R+RReQ\ϑ4yS1'l8hx ! \4yS^NirR+R(W4aglS5ul ! 9hnc4*NxW(W4aglSv~bkMOnQWV^R P$'f '3 'f'#5'f'f     #   3.2 Sf[MlKm^ W,gSt: (W [v8hxxW-N_eQSf[ƖV,Q(uSTirYt,ODNARP[(WOpvMOnM. N 3 $ff+b    b/g~$f  \SDNA7hTS:NUS ! k*NUSv TNeT+gz(u>e\'` TMO }h,NO>f:yDNAag&^ ! R+R(u N TelYt,_S]N*N8hxvMDNASO ! 5ul,SDNAv8hxz^ (Zy#     .Maxam-Gilbert l@b(uvSf[b/g "C (   Sf[lKm^[O$  3.3 ꁨRSKm^ G$fW,gSt N~bklKm^Stv T,S/f(u N TvgIQri_hddNTP,YddATPh~rgIQ,ddCTPh݄rgIQ, ddGTPhĞrgIQ, ddTTPh~rgIQ.1uNkyddNTP&^ gTyr[vgIQr, {S:N1u1*NlS Te$R4yxW.2~3c     3.4 ^8^ĉKm^ k~{5ul (uk~{5ulSNZNpQs^g5ul,we,R_Km^ۏ z,vQN z^ T~bklbSf[Km^l. 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OSRg OSRgoN(SignalP http://www.cbs.dtu.dk/services/signalP ) bKmǏ z-N[+T[temRNA 5 zvContigы:Nˆ}v^R; 6qT(uSignalPoN[MR50*N(lWx^R(N,{N*NATG[^v2ukx(lxMet_Y)ۏLċ0O YgSignalPRg~Qckb~g RKmՋ^R gS:NOS; GPY(WKmՋ^Rv,{N*NMet 5 zX[(W~bk[xP[ 勏^R:NOSvS'`f'Y0ZZ' >#  ' 3# # # # # ' f# ^     =- DB ~bk[xP[ ~bk[xP[: TAA, TAG,TGA GC% = 50% ~bk[xP[k 64 bpQsN!k GC% > 50% ~bk[xP[k100 200 bp QsN!k 1uNYpeWV ORF GWYN50*N[xP[ VdkgSv b^/f ORF N\N100 *N[xP[0 0PP!f   >0 C 3 zvnx 3 zvnx;N9hncPoly(A)>\^R,KmՋContig N+TPoly(A)^R R9hncR>\OS^R AATAAA TBLAST Tn'`k~gqQ T$Re0 b $f% 3% 38% 3% 3% 33   ?, zD ^x^R0Q+TP[ ؚI{w8huirYpeY>fP[^ N\N100 *N[xP[ gv N0R50*N[xP[uf\  > f1>"  @1 E [xP[OP1r'` x TN(lWxv N T[xP[y:N TIN[x vQ]+RN(W[xP[v,{3MOxW N T0 N Ty^\O(u TIN[xvs g_'Y]_ YN{|WV-N N(lxAle [xP[Y:NGCA,GCCbGCT, GCG_\O(u0$} fsh-.A2 NF Y>fP[ Q+TP[Lu Y>fP[TQ+TP[vLu gNNf>fvyr_ Y Q+TP[v5 zbyOSOMOdonor site 8^vz^:N 5 AG!GTTAAGT-3  3 zSySSOMOacceptor site), Y:N5 PyPyPyPyPyPyCAG-3 ( Py 'VvU8hx TbC)BP $fF#T1  B3 G N8nc6Rz^ QNN@b gWVbd~P[ N8n gc^R [NSNDNA~Tˆ}v\O(u c6RWVh0 SY*N+RuirvWV~yr g~b_NS\O:N$R+ROnc Y iRirWV~YWVv N8n gCpG\0 &zZtf8)@ C7 H oNKm Ǒ(uNCBIvORFKmoN ( ORF finder: http://www.ncbi.nlm.nih.gov/gorf/orfig.cgi )$ReORFvSV0HZfZ% 3 D6 B1.2 mRNAv5 zsSlU_wYMOp:S Ǐ Tn'`kegKmmRNAv5 z g8^(uvNlU_wYMOpvsQvpenc^/fw8h/TRP[penc^(The TRADAT Project , Eukaryotic Promoter Database, EPD. http://www.epd.unil.ch/ )0\ % % %% % q% DE4 1.3 Tng_ Ǐ]X[eQpenc^-NvWVz^N_gvWV~^RۏLk N-Ng~bSNKN9SMvxWz^SvQkO (uNLu[WVvely:N Tng0  ] fQYF5  Tn gY NQy`Q A DNA^RgNGrk[hQv T B _>exFhORF cR{|fP[ C _>exFhыb(lWx^RvvfP[ N T ONu}YQag^ NNvBgN&^ YgWV/fgNWV[evbXT_NOQsY*NOo` b Q~~NN'`TS6kv 4Z $ffr EH9 C WVhNir0N^v YgΘ^NO (ubNorthern BgNTRirBgNZoo-blotting Rg0 bNorthern BgN  9hnc]wvDNAz^_ir NmRNASO-NibXWVNir QNDNA:NcNKNBgN0Z     I: RirVBgN  9hncNsQ|vO?K<3.nx[DNAz^-NWVvMOn(f$ A Ǐ[hQcDNA^RvKm^0[k NSNWV~DNAvk nx[WV@b(Wv:SW B Ǐiry]^zW OVTirtVegnx[WVvMOnFFRr 'Vk;6rigT~mbS虨s t*0p56ihۭC1X3Nqc闶FE_<{-o{6s4O`+?ϧHz}>]LKUOs-C1׿*p;797ug P`Zc: e^͇Qz|*, Y{~ /Sw. 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OSRg OSRgoN(SignalP http://www.cbs.dtu.dk/services/signalP ) bKmǏ z-N[+T[temRNA 5 zvContigы:Nˆ}v^R; 6qT(uSignalPoN[MR50*N(lWx^R(N,{N*NATG[^v2ukx(lxMet_Y)ۏLċ0O YgSignalPRg~Qckb~g RKmՋ^R gS:NOS; GPY(WKmՋ^Rv,{N*NMet 5 zX[(W~bk[xP[ 勏^R:NOSvS'`f'Y0ZZ' >#  ' 3# # # # # ' f# T     =- DB ~bk[xP[ ~bk[xP[: TAA, TAG,TGA GC% = 50% ~bk[xP[k 64 bpQsN!k GC% > 50% ~bk[xP[k100 200 bp QsN!k 1uNYpeWV ORF GWYN50*N[xP[ VdkgSv b^/f ORF N\N100 *N[xP[0 0PP!f   >0 C 3 zvnx 3 zvnx;N9hncPoly(A)>\^R,KmՋContig N+TPoly(A)^R R9hncR>\OS^R AATAAA TBLAST Tn'`k~gqQ T$Re0 b $f% 3% 38% 3% 3% 33   ?, zD ^x^R0Q+TP[ ؚI{w8huirYpeY>fP[^ N\N100 *N[xP[ gv N0R50*N[xP[uf\  > f1>"  @1 E [xP[OP1r'` x TN(lWxv N T[xP[y:N TIN[x vQ]+RN(W[xP[v,{3MOxW N T0 N Ty^\O(u TIN[xvs g_'Y]_ YN{|WV-N N(lxAle [xP[Y:NGCA,GCCbGCT, GCG_\O(u0$} fsh-.A2 NF Y>fP[ Q+TP[Lu Y>fP[TQ+TP[vLu gNNf>fvyr_ Y Q+TP[v5 zbyOSOMOdonor site 8^vz^:N 5 AG!GTTAAGT-3  3 zSySSOMOacceptor site), Y:N5 PyPyPyPyPyPyCAG-3 ( Py 'VvU8hx TbC)BP $fF#T1  B3 G N8nc6Rz^ QNN@b gWVbd~P[ N8n gc^R [NSNDNA~Tˆ}v\O(u c6RWVh0 SY*N+RuirvWV~yr g~b_NS\O:N$R+ROnc Y iRirWV~YWVv N8n gCpG\0 &zZtf8)@ C7 H oNKm Ǒ(uNCBIvORFKmoN ( ORF finder: http://www.ncbi.nlm.nih.gov/gorf/orfig.cgi )$ReORFvSV0HZfZ% 3 D6 B1.2 mRNAv5 zsSlU_wYMOp:S Ǐ Tn'`kegKmmRNAv5 z g8^(uvNlU_wYMOpvsQvpenc^/fw8h/TRP[penc^(The TRADAT Project , Eukaryotic Promoter Database, EPD. http://www.epd.unil.ch/ )0\ % % %% % q% DE4 1.3 Tng_ Ǐ]X[eQpenc^-NvWVz^N_gvWV~^RۏLk N-Ng~bSNKN9SMvxWz^SvQkO (uNLu[WVvely:N Tng0  ] fQYF5  Tn gY NQy`Q A DNA^RgNGrk[hQv T B _>exFhORF cR{|fP[ C _>exFhыb(lWx^RvvfP[ N T ONu}YQag^ NNvBgN&^ YgWV/fgNWV[evbXT_NOQsY*NOo` b Q~~NN'`TS6kv 4Z $ffr EH9 C WVhNir0N^v YgΘ^NO (ubNorthern BgNTRirBgNZoo-blotting Rg0 bNorthern BgN  9hnc]wvDNAz^_ir NmRNASO-NibXWVNir QNDNA:NcNKNBgN0Z     I: RirVBgN  9hncNsQ|vO?K<3.nx[DNAz^-NWVvMOn(f$ A Ǐ[hQcDNA^RvKm^0[k NSNWV~DNAvk nx[WV@b(Wv:SW B Ǐiry]^zW OVTirtVegnx[WVvMOnFFRx: (: :XX9(  D  6~+  ,o 4͑SWV: TNkDNA :d&^$Ny N Tˆ}vvOo`. 40 f 6   0u+  M / ͑WV gN NQy`Q * N*NWV[hQ(WSN*NWVQ YBTA ETD vQx~gN Nv T d nF> nF nFJp       HD A"a  35P ( ,$D 0#z  (     ,$D 0W  TO+  ?? 8  7---ATG-----//------AATGCC ----//---ATAACG---//--TAA----x80CGf)CGfCC T 0   #  h B  ZD??0 0 B   ZD??  B   ZD??0  T 0    # l h B   ZD??0 0 B   ZD??  B  ZD??0  T 0   #   p h B  ZD??0 0 B  ZD??  B  ZD??0  T 0   # < h B  ZD??0 0 B  ZD??  B  ZD??0  T 0   #  h B  ZD??0 0 B  ZD??  B  ZD??0  T 0   #  lh B  ZD??0 0 B  ZD??  B  ZD??0    T\+  ??   RA*40GC 2   # l1?#" `L ( 2 ! # l1?#" ` l( 2 " # l1?#" `l ( 2 # # l1?#" ` ( 2 $ # l1?#" ` ( 2 % # l1?#" ` ( 2 & # l1?#" ` <( 2 ' # l1?#" `< ( 2 ( # l1?#" ` \ ( 2 ) # l1?#" `\ ( 2 * # l1?#" ` | ( 2 + 3 r1?"| ( 2 , # l1?#" `| ( 2 - # l1?#" ` ( 2 . # l1?#" ` , ( 2 / # l1?#" `, ( 2 0 # l1?#" ` L ( 2 1 # l1?#" `L ( 2 2 # l1?#" ` l( 2 3 # l1?#" `l ( 2 4 # l1?#" ` ( 2 5 # l1?#" ` ( 2 6 # l1?#" ` ( 2 7 # l1?#" ` <( 2 8 # l1?#" `< ( 2 9 # l1?#" ` \( 2 : # l1?#" `\ ( 2 ; # l1?#" ` |( z `@  < B,$D 0T 0   =# 0 B > ZD??0 0 B ? ZD??  B @ ZD??0  T 0   A# 3 0 B B ZD??0 0 B C ZD??  B D ZD??0  T 0   E# ^0B F ZD??0 0 B G ZD??  B H ZD??0  T 0   I#  0B J ZD??0 0 B K ZD??  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