JP 2009-249304 A 2009.10.29 10 (57)【要約】 【課題】乳癌分子標的
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5. 10000000 100000000000000000 000000000 10000000000 00 0 7000000000000 100 720000 1000000000000000000000000000000 00 0 100000 2
6. 1000000000000000000000000 000000000 10000000000000000000000000000000000 000 0000000000 ID HH 2 LU UL 010 500000 10 300 0 11100 0000000
7. 000000 BCcol 00000000000000000000000 00 00000000000 O0 O0 0 O0 100000000000000000000000000000 0 0 0 01 100000000000000000000000000000 Pratt Transcription and Tranlation Hanes D Higgins S J IRL Press Oxford 179 209 1984 00 0000000000 coli 530 extract system
8. N m O _ MEE t ug s E rs pp S ELS s L3 Ep qe t n 3 2 r3 L1 Merken _ O _ gl C3 _ 0 r3 L3 r3 ss mm 1 L3 r3 fs t Nm N C sx m x EI ET g Expo _ _ mu ENES am iN en aduer Q 3 HL L3 r3 L3 r3 8 Bobo BD lt rS Q 3 3 Q 3 3 r3 3 e L3 r3 L3 nm m 8 o a L3
9. tu uuu 100 10000000000 00 0 263 267 1995 Virology 0 520 456 19730 000000000000000 jiululudgiuiuiluildiuiliuluslululkulul 1000000000000000000000 500 20 U 0 l RPM 1640 1991 s F 120 0000000000000 p000 6008 30 49 150 H 60000 0 0 0 0 0 0000000000 8 HM gt
10. 10 P 2009 249304 2009 10 29 000000 0 0 PCR 0 DNA BCG01 DNA DNA 100000 0000000 00 00000 Molecular Cl oni ng 000000 100000 0000000 000000 00 00000 000000000000000 Mtan super Express U DD DD D 00000000000000000 004 Gapped duple x0 0 0 000000000000000000000000000000000 100 5440 0000000000000000000000000000 10000000000000000000 240 005 0000000 00 000003 0000000000000 0 0 0 O O TA 900000000000000000000000000000000000 00000000000 1000000 BCG01 0 2940000000 5440 00000000 9000000000000 00 00000000000000000000000 2883220 pBR3250 pUC12 pUC13 000 110 pTP50 sHi9gj 5 15 0 0 pFast Bac HH 1 11 pRc pRc RSV pc DNAI Neo
11. 000 000 10000010000000000000000 Bodanszky 000000000 0 Peptide Synthesis ntersci ence Publishers New York 19660 Schroeder O Luebke O 00 0 O O The Peptide Press New Yor k 19650 10000000 10 0 000 10000000000000000000000000000 10000000000000 DNA O0 O0 O0 00000000000 DNA 1000000 DNAD O 0 0 O DNA DNA 100000 500000 00000 1000000000000000 00000 1000000000000000 DNA 0 0 0 00 lt
12. 000000 Fw 5 CAAATGGTGTGAACCCAGGG 3 Rv 5 GAGTTGGGAGGTCAGGGAAGA 3 100 14000 D D 00000 wg O 5 3 O0 000000 5 TGCAGGCAGTGCTCCTGAC 3 000000 HBL 1000 McF 70 MDA MB 4530 OCUB FO OCUB SK BR 30 T47 D ZR 7 5 10 ZR 75 300 D 0 B 0000000000000 Aneric a Culture Collection Manasas 000 0 000 0 0000000000
13. 1000000000 0 0 0 100 Fatel serum Equi tech Bi o I nc l 37 50 10 0090000001000 1000 10000000000000 000 000000 si BCO 1 1 20 00000000000 0 control si RNA MCF 7 72 RNAU 00000200000 0 0 151 strand cDNA doub le strand 00 0 0 amplified aRNA 0 0 si Cy5D 000 aRNA 100 161100000 000000000 utt 000 100000000 0000 000 15000000000 10 1000000000 200 0000000000000 10 20 0000000000040000 10 1000000000 000000 0 O0 ast Cancer Genes 10 01 si RNAD O0 0O O 0 O steal th si RNA i nvitrogen Bcc oip o0 000000 20 si Anaxap 0 000000000 00 0 00 0 0 0 0 0 0 0 0 0 cF 70 00000 0 0 00 0000 00 00000 000000 000 20 2009 249304 2009 10 29
14. o o o N r3 o r3 3 E zi Eb EP ER e r3 3 r3 O E 3 3 Ex 3 i E 5 er Me uq L3 3 3 r3 zi 525 3 O 3 C r3 r0 r3 3 L3 m lt s gt r3 3 L3 e gt ux EN r3 1 3 p ME ix II gt x o r3 r3 m r3 gt D E r3 ma r3 r3 g 3 r3 a v L3 r3 3 r3 m d I 2 ES EUER 3 Cj 1 r3 r3 3 s p 3 3 3 3 3 PENES Run lt 3 L3 r3 3 Lj 3 L3 3 _ r3 L3 r3 r3 ET r3 _ r3 3 L3
15. 00000 100000000 _ 70 86601000000 U siRN 00000000 51 j D 720 D 10000000000000000000000000 50 10300000 5 24 well Cell Invasion assay CELLBI OLABS CA USA Doubl e chanber si RNAD 0 0 0 0 McF 70000240000filter000000000000000 M LAT 10 0000 1500000000000000000000000000000 43 150000000000000000000000000000000000 20 00 20 0000 400000000000000000000000000000000000 23 20 000000 Hu Breast Cancer 01 BCGO 1 000000000000000 0000 00000 BCG 1 10000000000000000 0 22 155 strand BCG010 0 0 0 100200000000000000 1000 HE 000000 30 1000 000 0 000000 o 00000000000000 00000000000 0 0000 1st strand cDNA 100000000000000 70000000000000 00000
16. N in 3 3 3 C3 C LJ L3 C L3 C O r C NDS 8 ES Em O _ 3 3 3 3 t m 2 3 31 3 YI TY gt psi E 3 3 3 3 o E EE 5 C3 3 E 3 3 3 3 s c E 9 oo 3 3 3 3 3 3 o I s e EN 3 3 3 3 3 3 ea E E 3 3 3 em 3 ec 3 2 C3 m aporte ES C3 r3 2 rrt m C3 C3 m am m m 3 3 Ep m m m s O 3 3 3 f r3 r3 3 3 3 3 LJ LJ 3 3 m IEEE E E st 3 n L3 3 3 3 m m m o 3 3 3 3 Bend d LAT Mtm E MI
17. T i nvi torogen 5 dNTP M xture Takara O IO 400 RNasin RNase hi bitor Promega 2u I 10pm gt 57 3 6000000000000 0 0 0 0 50410000 420 00 0000 0 0 lt 1000 10 000 10 10000 RT PCR O st strand O0 O0 0 1 10 0000000 0 0 0 SYBR Green Master 140p 10 ABI 7900HT Sequence System 1 50 C 20 2 95 100 3 95 15 4 60 3 0 0 0 00 400 0000 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 11000 000 5 GGCGGATGICTTGGATAGATCA 3 010000000 5 3 10000000 BCG01 2 0000 5 3 00000000 5 3 00000000 ll 1000000 RI bufferD 2 5
18. 2009 249304 2009 10 29 13 BCGO1
19. 4 2009 249304 2009 10 29 UJ HM UJ 10 20 30 0 5 2009 249304 2009 10 29
20. 1 0 0 0 000000000 0000000000000 BCGOIU o0 0000000000000 00000000 0 0000000000 80 90 95 970 0 98 000 00 0 0 00000 0 00 457700000 Lys Arg His 00000000 Gul Asp 0000 00 00 00 0 ser IDD 00000000 0 Thro 0000 10 100000000 10000000 100010100 000 10000000 10 10000000 10 000 0 Bow 0 Sci ence 2471 1306 1310 1990 10 0090000001000 100000000 BLAST Nati Center for Biotechnology Information Basic Local Ali gnment Search Tool 00 0 000000000000 8060 620 101000000 OFF 0 00 00090000000000000 0000000000000 D D Karling Proc Natl Acad Sci USA 900 5873 5877 1993 000000000 000000 00 0 0 NBLASTO O O 0 0 O O O version 2 2 00000000 Nucleic Aci ds Res 250 3389 3402 1997 0 Need J Mol Biol 480 444 453 19701 000000000000 100000000000 10000000000 Wersi Mller 05 4 11 17 1988 100000000000 ese 1000000000 ALIGN 00 0 0 version 2 0 Pearson Proc Natl Acad Sci USA 85 2444 2448 19 88 0
21. P P e gt 1 control siBCGO01 1 siBCG01 2 siBCG01 1 2 400 350 300 250 200 T 150 100 50 0 1 2 1 2 cont IL 6 13 0 00003 0 0003 0 000025 0 00025 0 00092 0 0002 0 000015 0 00015 0 00001 0 0001 0 000005 0 00005 1 2 1 2 cont 971 2 1 2 cont 0 0014 RAS guanyl releasing protein 1 0 00035 RAS dexamethasone induced 1 0 0012 0 0003 0 001 0 00025 0 0008 0 0002 0 0006 0 00015 0 0004 0 0001 0 0002 0 00005 071 2 142 cont 971 2 12 cont 0 00006 RasGEF domain family member 1A 0 0014 pregnancy specific beta 1 glycoprotein 6 0 00005 0 0012 0 00004 0 001 0 00003 0 0006 0 00002 0 0004 0 00001 0 0002 0 0 1 2 1 2 cont 1 2 1 2 cont 0 0008 Pregnancy specific beta 1 glycoprotein 8 0 0008 Pregnancy specific beta L glycoprotein 9 0 0007 0 0007 0 0006 0 0006 0 0005 0 0005 0 0004 0 0004 0 0003 0 0003 0 0002 00 0002 0 0001 0 0001 0 1 2 1 2 cont 1 2 1 2 cont Snail 2 TGF heta2 0 00014 0 0012 0 00012 0 001 0 0001 0 0008 0 00008 0 0006 0 00006 0 0004 0 00004 0 00002 0 002 0 1 2 142 1 2 1 2 cont cont 27 interferon induced with helicase C domain 1 interferon induced protein 35 0 0007 0 0006 0 0005 0
22. UJ 3 HM a 01 b BCG01 10000000000000000000000000 00000000000000 17 2009 249304 2009 10 29 BCG01 3 CDS 000000000 BCGOI 3 1000 BCG01 4 CDS 00000000 BCG01 4 1000 10000
23. 0010 2009 249304 2009 10 29 3
24. 570 000 100000 100000000000000000000000000000 0 00000000000000000000000000000000000000 10 Bc 10 000 100000000 10000000000000 100000000000000000000000000000 Bc 1 0 0o 000000000000000 100000000000000 uuu D00000000000000000000 0 10 2009 249304 2009 10 29 2 10000000 n 10 RNA 20 30 1000 40 1 50 l
25. 10 20 30 40 50 22 2009 249304 2009 10 29 62 00000000000000000 100000 4 10000000000000 70 00000 si 72 si O0 0 0 0 0 0000000000000 30 si Cy 50 0 0 0 0 00000001060 1000 16 5440000000000 1100000000BCG0000000000 1040
26. 2009 249304 2009 10 29 7 30 40
27. 5 w Se P Kc 5 amp 5 X k kJ 3 93 S o 5 x0 8 5 e 0 01 0 0018 0 0016 0 0014 rH 0 0012 I 0 001 0 0008 9 0006 9 0004 0 0002 G 4 Ra e lt lt lt BCGO0I 2primer GAPDH 0 0014 0 0012 ri 0 001 0 0008 0 0006 0 0004 0 0002 Control SiBG01 1 lt cadherin18 siBG01 2 siBG01 142 P cadherin VE cadherin siBG01 142 Control siBG01 1 5 01 2 lt gt 4 cytokeratin14 lt 18 gt PCNA amans N cadherin cdkl cdc2 md ou 53 e psp Supe cdki e a Gs cyclin w gp qua quae lt 7Jv2Z2 RT PCR gt E cadherin cyclin mur 2500 0000 cyclin D 2000 0000 1500 0000 cyclin 1000 0000 500 0000 n p21wafl 0 1 2 1 2 cont tubulin JP 2009 249304 A 2009 10 29
28. 100 BCG01 i 5 RT PCRO 00000000 2009 249304 2009 10 29 15
29. O Q HM HM E E E HM Bur 00000 00000000000000000000 51 O00000000000000 01 si 0 7 01 BCG
30. OUUU 00000 10000 0000 MP 130 IL 60 000000 13 000000000 0000000000 0000 10000000 1000000000000 0000000 0 10000000 11 600 0 0 0 0 0 0 13100
31. DNA O0 O0 O0 0O O mRNA 1000000000 Chai n Reacti PCR 00000 Reverse Transcri ptase RT PCRD LU Ul 10000000000 J DNAD O 9544000000000 cDNA B 10000000000000000 U UUJUYLBBUBgilubltuu ul LU UULHBUYEBUBBUBdiWulbWubiulbuliluiluiliuib illiuil 10 20 30 40 50 9 2009 249304 2009 10 29 00000 DNA 000 90000000 0 um 000 100 1000 00 DNA DUO0O000000000000000000 80 100 90 95 1000 97000000000 980000 1000000 DNA 100 100000000000000000000 NCBI BLAST Nati Center for Biotechnology Information Basic Local Ali gnment Search Too 00 00000000000
32. O06 GO9 CAD GN8 H8 9 HN2 HA7 48063 018 18 WO Q3 32 36 qo5 862 Q69 77 882 616 625 634 636 002 Aso2 6 1 BA22 28262
33. 0 1M Tris HCI 8 12 2 mercaptoethanol 200 9 Bromoph 18 2009 249304 2009 10 29 SDS pol yacrylal de gel 10000000 BST 20mM Tris HC pH 5 100 0 x 150mM NaCl 10 4 16 100 00000 ECL Pl us V stern Blotting Detecti System healt dNTPs interfer
34. RTS 500 Rapid Tranl ati on System 0 000 10000000 Rabbit culocyte Lysate System Promega 05 0 00000000 100000000000000000000000000000 Johnston Nature 179 160 161 1957 Eri ckson Meth Enzymol 96 38 50 1996 J lU 00 0 0 00 00 0 lt Pratt J M J 1984 J H tP P TD DD BD BD T BUT PEDO Spirin A S Science 242 1162 1164 1988 0 00000 10002 11000000000 060 00000000 0 PROTEIOS Wheat pro tein synthesis core kit 0 0 0 0 O0 1000000000 2000 3336730 000000000000 MP13 18 10000000000 00000000000000000000000 000000 000000 1 BCGO1 0000000 1000 10000000 000 Bcc oigp 00000000 10000000
35. 1000 BCGOI1 N 3 oo 3 3 lono 3 r3 3 3 L3 3 zs EE Iz un AMEND 5 Te 3 3 3 E 3 N 3 3 L3 3 5 Im _ N N E Q L3 O L3 Dj 3 3 zi Di EE 3 L3 3 3 3 3 r3 3 _ _ _ 3 25 H t _ r3 3 3 Im Q 3 3 3 __ 1 3 3 O 3 3 3 3 Lj ER I m 3 O 3 3 3 3 I __ m O 3 3 3 3 L3 3 E EJ d E _ E un lt 3 3 L3 3 3 o 3 3 3 3 3 3 3 o g 3 L3 r3 m OS OMEN E EXEC xx X ACE LE nd
36. HM BCG01 1000000 0 5 2096 1000000000000 RPM 16400 199 12 0 000000000000000000000000 00000 0 1nM 100nM 10 24 _ s RE 0
37. 66 5 00009000 1000000000000000000 000000000 170000000 1100000000000 11 60 0000000000000 130 RAS nduced 1 5 guanyl easi ng protein 1 calci um and DAG regul RasGEF domain member pregnancy specific beta 1 gl ycoprotein 6 8 9 TGF 8 2000 000 epithelia mesenchymal transition 1 2 Sl ug 1000000 100000 interferon induced th helicase C domain 10 i nt 1 nduced protein tetratri copepti de repeats interferon al ible protein 60 i nterferon i nduced protein 35 interferon al pha i nduci bl e prote in 27 i nterferon sti ated transcription factor 3 gama 48 CXCL14 00000 100000000000000000000000 00000 00000
38. O _ Q 3 L3 3 L3 r3 3 3 Td _ oO rx i edd 3 9 EN Umm 5 C e oog L3 r3 m 3 L3 3 I O _ m DEI E 3 I ES EX em ELS hl lc o L3 L3 m a E us EE E E Q L3 E t 3 L3 Lr _ r3 d 1 1 AMNES _ L3 E 61 224 ET El EX 5 Q 3 O m m 3 3 3 3 L3 C a ms m _ REN n d pap 8 Q r3 o a5 Lr O I _ Q e Q RE N Q s 5 m ac 16 2009 249304 2009 10 29 0000
39. 0 0 00000 10000 10000 BCGOl D 0O 00 DNA 1000000000000 1 00001 RefSeq Accessi on 0209740 0000000 BCGO01 J 0002 000000000 010 000040000 000000000600 000000 GenBank Ref Seq Access on No NM 020052 1000000 10000 1000000 10000 1000000 0000 0 0 0 000000000 0 BLASTI FASTA 0 D Jackson Muse Genome Informa ti cs http www informati accession Othologyp 0000000 100000000 0 0 00000000000000000000 10000000000000000000000 000 0 0 10 0 iupdgbiugdudtiullululiuull li 1000000000000000000 BCG010 100000 0900000 00000000 00000000000
40. A n A 3 EE m i 3 3 3 ETE nz t IX C3 3 Cj g 3 3 3 3 3 mm _ 5 m 3 3 m toe AI ere aie em ne 3 3 3 o C3 n e 3 3 C ___ L3 LLI O 3 3 3 rar 3 Ed 3 3 _ 3 L3 r3 3 L3 PN ux m GUN m 3 3 C3 3 O m m L3 r3 r3 Ej _ em lt 2 A L3 pd o 3 3 a L3 3 C 3 r3 t 3 m 8 222 Im 3 3 3 Q g gt s C g C3 C3 pu C uw 3 G m X AE O 3 3 3 es LI 2 L3 cm m LI c Q 3 3 o r3
41. 1 2 HM CJ 12 2009 249304 2009 10 29
42. 1000000000000000 BCGOID 00000000000000000000 0000000000000 70 00000 720 00 00 00000000000 1000000 240 0600 000000000000 0060 23 3 28 60 si BCGO1 10 101 0 16 8 si 01 20 159 3 62 20 si BCG01 122 296 0 40 60 U U DD D D D 100000 100000 00000 100000 0 0 0 MCF 7000000 000 100000 00000000 10000000000 10000000000 0001000 3 010 1000000000000000000 1000 1000000000000000000 10000000000 si 0 7208 000000000 00 0 siRNA 14 N 10000000000 00 14 18 0 0 1 cdc20 i nA 53 1 H 0 0 0 O0 10000000000 1000000000000000000 10000000000 E gt g a E E T A 000 10000000000 cadheri 180 BCG01 BCG01 10000 100 P H 000 100 10000 10000 1000 10000000000000000000 BCGOL 10000 cycl i nA 1 cdc2 BCG01
43. 3 5 3 one i nduced 110000 Fw 5 3 000 Rv 5 AGATAATGCCCCGTGGGTC 3 000 amly menber 1A400000 Fw 5 GCTGTACCATAATGCAGGACGTC 3 Rv 5 3 5 TGTCTGGTITGCCATTGITGAAC 3 5 GCACTACAGGTAATCAAAAAAAGGC 3 Fw 5 TGGAGCATGCCCGTATTTATG 3 Rv 5 AGGACCCTGCTGTGCTGAGT 3 10 20 30 40 50 19 2009 249304 2009 10 29 000070000 0 5 3 Rv 5 3 i nterf eron i nduced protein 350 0000 0 FW 5 GGTAGAGGCCCTGACAGTCG 3 00000000 Rv 5 AAGACTGCTAGGCCCTGCTG 3 0000 Interferon al pha i nduci bl e protein 60 0000 5 AGAATGCGGGTAAGGATGCA 3 0000000 5 TGICCGAGCTCTCCGAGC 3 i nterf eron i nduced protein wth tetratri copepti de repea 0 FW 5 CGGAGAAAGGCATTAGATCTGG 3 uuu Rv 5 3 interferon al pha i nducible protein 2710000 0 FW 5 CAGTCACTGGGAGCAACTGGA 3 00000 5 GCCCAGGATGAACTTGGTCA 3 i nterf eron sti ated transcription factor 3 gama 4
44. BmNPM AcNP MESH gt 0nnnnnnnn pe HM TOR SRa SV40 LTRI 0000000000000000 0 0 85 qr 00000000 LTR HSV TKI 000000000 100000 0000 00 SR 10000 1000000000 SV40 SV40 ori 00 000000000000000000000000000 10 20 30 40 50 11 2009 249304 2009 10 29 0 0 000 00000 EP 00000 COS 7 Vero B u 00000 000000 9 CHO 0 0000 At T 200 2930 10000000000000
45. 3 un O I O __ Em lug __ _ O I E Q 3 3 m vp O I EC L3 o g 3 3 O I Ej 3 3 O I 5 Q gt m m O I _ N O I 50 0000 BCGO1 2009 249304 2009 10 29
46. 00000000 00000000 00 0000 10000001000 10000 10000080100 900 00000 95 9770100000000 98 10000 0 O0 O0 0000000000000000000000000 utt 10 20 30 40 50 6 2009 249304 2009 10 29
47. 19 51 Int Cl A61K 38 0 A61P 35 0 C12Q 1 68 120 1 0 GO1N 33 50 21 HERE E 22 H B H 2006 01 2006 01 2006 01 2006 01 2006 01 882008 96639 P2008 96639 20 4 2 2008 4 2 19 FI D 12 OL 54 RAOR k ORBE AGIK AGIP 120 1 68 C12Q 1 02 GOIN 33 50 31 02 35 00 aF 98 A 74 72 72 2009 249304 2009 10 29 11 2009 249304 2009 2493044 43 21 10 29 2009 10 29 2 45 48024 48063 40084 ZNA Z 28 504137912 J 3 1 110000154 A i 2 T SEES P 3 1 J 3 1 3 20045 AA34 5 802 DAI3 DA36 lt
48. 00 000000000000000 40 100000000000000000000000000000 1 BCG01 1000000000000000 BCG01 70 D 700 00 0 0 8690100000 1000000000000000000000000000000 8 000000 7 0000000000000000000000000000000B short interfering si O 0 si BCGO1 10 5 UCUUGUGACCUGAG CUGCAUCGUAA 3 0 0 0 0 0 0 0 O si BCG01 20 5 GGAGAUUUCACUGGGUACAUUGAAU 3 si 01 1 2 mxture0 0 0 control si 5 UCUAGUUCCGAGGUCUACG CUGUAA 3 000000000 7000 000 00 siRNA 0 0 86601000000 50 2009 249304 2009 10 29
49. 0000 100000 300000000000000000000000000000000 000 1000000000 Mol ar J 5 et al Cold Spri ng Harbor Lab Press 1989 0 10000 6x 55 sodi um chl de sodi um ci trate 0 0 45 0 0 0 0 0 2 5520 0 1j SDS 6 10000
50. 00000000 10000000000 ECM 10000000000 1000000000000000 2 IEEE T 79 E rm UJ 2 HM
51. 00000000000000 00000 BCG010 1000000 000000 100000000 Rasg 0o 000000 0 0 0 RasGAP RasGEP 0000 Ras 100000000000000 100000000000000 1 pregnancy specific beta 1 gl ycoprotei n PSG 10000 pregnancy specific beta 1 gl n O O syncyti otrophobl ast 100000000000000 1000000000000 0 0 9 0 0 anti gen 0000000000000 00 000 1000000000 8 010 l PSG 0000000000000 0000 70 00 000000000 0000000 23 2009 249304 2009 10 29 Snai 0 1 O EL N CG UJ 2 HM UJ
52. 00000000000000000000000 E cadherin VE cadherin P cadheri N cadherin PCNA cyclin cyclin n E 21 1 1 2 cdk2 BD transduction USA cyclin 1 BD Pharm ngen USA tubulin p27ki p1 Cell Si gnali ng technology UK cytoke rati n14 cytokeratin 18 Progen USA cadheri n 18 Abnova Anti rabbit IgG HRP Li nked Cell Si gnali ng Anti mouse IgG HRP inked Cell Si gnal i ng technology 0000000000000000000000 10000 151 strand total O O Nano Drop 1000 Nano Drop Technol ogies RNA 0000 25 90 RN 0 0 0 0 0 0 D 0 D 210 08 0 5901 0000 2u 10 200U u Supers cript 11 invitrogen 10u 5x First Strand Buffer invitorgen 10 0 1 DT
53. 00001 50 30 200000 ii 1000 1000001000020 10 50000000000 10 300 0 0000 10 10000000000 50 40 3 20 0000 1 2 11 50 100000 30 10 100 0 000100050 40 300 0 0 2 JJ U U UU lU 10 500 0 10 300 0000000000 100 1000 50 40 30 0 200000000000000000 l Ul v U UUUYLHBBUBubgubuuib u u lu 10000000000000000000000000000 1000000000000000 CB j jl 10000000000000 BCG0O1I0 Refseq Access NP 0660250 00000000000 0 8690110000 2000000000 00003 UJU JU H H 1 00004 Ul 10000000 GenBank RefSeq Accession NP 06 1 LU UL 000000000 10000000 LU UL JU JU H tL LU Dll BCG010 C 1000000000 0909000000 CONH 10000 COOR 100000000 UULHULLUt 9091091090 0000 1 60 0000 3 80 0 0000000000 0 6 12 0 1 200 FL U 000 00 7 14 LU UULHLULHLEULHUILIULD U 0 0 9 DBiulbiluilulkilisisu lll
54. 0004 0 0003 0 0002 0 0001 0 9 00009 0 00018 0 00016 0 00014 0 00012 0 0001 0 00008 1 0 00006 0 00004 m 0 00002 a 1 2 1 2 1 2 0 1 2 cont cont interferon induced protein Interferon alpha inducible protein 6 with tetratricopeptide repeats 1 0 00008 0 00007 0 00006 0 00005 0 00004 0 00003 9 00002 0 00001 0 0 0016 0 0014 40 0012 0 001 0 0008 9 0006 E B 0 0004 0 0002 LM 1 2 1 2 1 2 interferon alpha inducible protein 27 cont 1 2 cont interferon stimulated transcription factor 3 0 01 gamma 4 0 008 0 007 0 006 0 005 0 004 0 003 0 002 0 001 0 009 0 008 0 007 0 006 0 005 0 004 0 003 B H 0 001 k 0 0 00007 2 122 cont 1 2 1 2 cont CXCL 14 0 00006 0 00005 0 00004 0 00003 0 00002 0 00001 aM 0 2 1 2 cont 2009249304000001 app 2009 249304 2009 10 29 28 2009 249304 2009 10 29 1000000000 51 11 1000000000 0000 3915 2006 01 3315 1509 2006 01 0000 15 00 1000 48024 01
55. 01 E 100000000 000000 01 01 10000 24 2009 249304 2009 10 29 10 20 30 Bh 01 2 E 75A5 amp 7525 8 C SEX LES 1 014 012 0 1 0 08 0 06 0 04 0 02 25 2009 249304 2009 10 29
56. on induced wth helicase C domain 1 5x buffer 10u 100000 5 101 I 2 IO SQ 5 5 Phusi 10 98 300 000000 00 98 C0 100 10000 0000000000100 0000000000000 000000 Fw 5 AGTGICCCCCGGTATCTTCC 3 000 Rv 5 ATTTCCAATTTCATCGGGATTG 3 5 TGGCAGAAAACAACCTGAACC 3 Rv 5 AGCATCCATCTTTTTCAGCCAT 3 0D B l Fw 5 GATCGGCCATCAAGGGAAG 3 Rv 5 3 ific beta 1 gl ycoprotei n 6 0000 5 CGGGCTCTATGCTTGCTCTG 3 Rv 5 GAGATTTCCTTGCCAGTGGCT 3 ific beta 1 gl ycoprotei n 8 0000 5 TAGCGGGCTCTATGCTTGCT 3 0000 Rv 5 CITICCTTGCCAGIGGCTG 3 000 ific beta 1 gl ycoprotei n 9 0000 5 3 00 5 TCCAAAAATACTCTGCCGGTG 3 easi ng protein 1 0000 FW 5
57. r3 r3 3 3 3 E r3 Ri 28 r3 g g r3 Q O Lm r3 i 3 3 r3 E nm e B A 1 O O 3 3 o 3 r3 O O 52 I O 0 L3 Lj 3 C3 3 3 L3 r3 ro EIE m ACT E 5 ux m LI N _ Hs _ ro ge 8 r3 Q t O I 3 8 CI E x t s ENE 9 Im 5 3 Q 14 2009 249304 2009 10 29 L1 UJ 3 HM
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