(19) EA SEAN RRA (12) RASA A ARAB [51] ine. cr” C129 1/68 C12Q 1/34 GOIN 21764 [21] HHH 998051519 (43]8%8 2001 4F 59 308 [11] agrs CN 12974878 (22]@MB 1999.3.5 [21] Ry 99805151.9 [30] ¢eaea8 [32]1998.3.6 [33] DK [31]0308/1998 [86] RRA PCT/DK99/00112 1999.3.5 [87] MRR © WO099/45143 BE 1999.9. 10 [8S]. miaRBRBLEM 2000. 10.17 (A PRICE sent FRU (72]RMA L- BES K+ S + FPR J+ RARER C+ T+ SPREE Ts Ls SIRES + ERSTE Sh [74] Se PRR REE eA SR A RAE ABR 6 RG 43 HT RT [se] ema SOAR TRO (57) 9m SRE A a He Be ERE, ETT OWES EOLA MATER MIE MR, RBA WALA WE A RE IY RS RE i. 1SSN1008-4274 BOAR F AR HAL BHRRAZR FH 1. i 3&2 AE AOS Oh ERK, RA OE: alERKAAPREGAERFA PEA MOM, b) RAMS 5 HS KAISHA RB, c) RE A HY REAR, fo d) BER AMRAA RGM RERB. 2 BAER LOA, KP RERRRM IMR AE / RAR, BE GPA LE ART RAROHRALH. 3B. RARRIMG HR, RPRAERRAMBRERK. 4 BRARZRIAFTA, EY MBAFAD ae. 5. BALK 3 HAR, Kp sie Re MISH, Rp eT SAR AREBREORALH. 6 RALKR ARS HH, RPMHRARALRA MB, & 2 By bm Fo JAK 3 WL 3H. T.BAERGE MAK, KPMRMBAFRH AHA. BBAERKR 6 HAH, LPARBRAARB MB, ABA 4h ba Ke, 9 RARK 5 HHA, LPRIRRHABLKASHAMMB Hp ih Sa. 10. RA RR 5 WHR, HPMERAARAAA DNA, RNA, cDNA PALI B HE BFA KP LL RAKRK 10 HAR, KP RIM SR RM AB, MEF PCR MRAM HSH. 1A RABRSHAHE, RPBLWRAAwH, FCEWPAH. 13. KARR 12 MAH, HP RR EMRE LSRE RRA RICS MD. 14. BALK 13 WHK, HP RAR BLMOLFRLE AEDN OBE BARR. 15. RAZR IZHFK, PRLMBAAMH MH HH. 16. RAKK IS HHA, PAM RAMA SJ2. 17. BARAK 12 HAH, KEP FH EW ElectroMAX DH10B 4 fe, 18. BAEZRIZAFTE, RP REMBAF HERA HH. 19. RAZR IHFHE, PARARBHR. 20. RABLK 12-19 HH, Kp aE aA ALT. 21. RALHK 20 HAH, HP MA pSJ1678 XK pZEr0-2. 22. RAKK 20 WF, PRBS RT 10 eS AEREARE RFE. 23, RAIRK 22 WHR, HP AH eZ eK A Fl RARARE: REE, OKA, FRE, AFHSA, SEHK zeocin. 24, LEE-RAZRAAK, EPHRABLEMRAEGR, B Mik Fa hik, 25. RAZR 4A, HPSWLSw EM. 26. RA KK 25 HAR, HP Be A AGE RB (EC 1.-.-.-), 445% (EC 2.-.-. -), RAPRE(EC 3.-.-.-), ROME(EC 4.-.-.-), Ft Aa (EC 5. -. -. -) Fok 4R ME (EC 6.-. -.-). 27. BARK 26 WH K, KP RAE A AAA, PRA, BK , RKB, ADDR. RRS, Bee. 28. RA BK 24 HFK, RPAWLEWAARAWALABH 4A. 29. BAER 28 HFK, EPSWRSWHATALAMAKY Bh LMR. 30. KARR 29 GFK, LPEWKSWAME. 31. LEB-BAERAG A, RP RAWKARK AML HID HAMIT. 32. RAZR IL HHH, HPAMATFRS RA, FRKAREK. 233. RAZR 31 HFK, RPRAMLW MET AMA FE 0. 01-10% 7 BEE SHH EB. 34. RAK 31 HFK, RPRAMAHMUAM RM. BHR ARHAMA FAS. 35. RA LK 32AFH, RPHMAFRARAM, Kh, REG, MEO, $48, He, WH SRA R. 36. KARR 35 HHH, RPRGOMRKABEG, MHA wy, WK, GEA, KZEA, FREER, PLERK, ARAE GRAWEKEA. 37. BAEK 35 HHA, EPRAP I. 38. RALK 35 HF, RPSRRARPDRR, RR/FRE LEER, FREFPEARE. 39. RAZK 38 HHH, RPSRRAAM RR, ARH, BE 48 i RAB Fe M4318 RH, PASC, RFLBRR, UHR, AMR, LRH, BRA. 40. RAZR 31 HF, RPAMAT TA ADM, A, HHA MADER. AL RAZR 410 HHA, LPHMA FM FR BRB HK BL, 42. RAZR IL GFR, LPHAMATFAKFABRSRM. 43. BAER IL HGH, EPRAMLHAF HEALS. 44. RAZR 31 HAR, HEP RAMA 350-700nm TH AA RKB, 45. RAZR 14 HHH, EP RAMIEWA 400-600nm HHA RAMA. 46. RA KK 31 HAR, EP RAMI 400-800nm ZH AA BRKRMH. AT. BARR 46 HAR, HP RAAIZH HE 450-700nm VHA A RKAH. 48. BARK 31 HFK, HPRAMAZHHAMATPRERA 3ARID Z Wel oh Z 1 SEH A 20-50 HK. 19. RAZR 31 HAH, HP RAMDHWHRLRRH AA 1-7 we. 50. RAIKK 31 WH, HP RA HK 0 tee ALI | A 50-500nP. 51. RAZR 31 HFK, IP KAMIWIK A BODIPY AM, K KE, FRA, ARM, CLPMAARRERA. 52. RARKIZGGA, GUHR: a) HSRALH, D)HLFEHRARHPFAERAMES P, oo) THD HORA Hw, d) IMEI R/ BET HG BH BA AIK HM, EERE LERPHEMKEDREKEH RS RADAR LEEKS HE RAW H-RRB, g) 5 ES BR BER RH HH PP RAE. 53. RARER IDK, GUT R: a) FE Si fo AES A RE BF FI 04 SH, b) HWRF FAKE MEE YP, c) FG DB th B01 AH do BS Ok BBP ES Se d) SEARLES P Sol AH AAR A, ) 4 RAW K-RRE, £) Bi 4 WBA WIE DRS AER KH Kh oy FP RAL. 54. BARK 53 HHH, MPH MR a) RH AE SAH eT 1-5 4%. 55. BARR 53 WHA, LP HR b) MU HH RHEE Ss DPRELA 0.5-10 MEH F/LEH FP RHR ERASE PR BET. 56. RALR 55 HH, HEH b) MUHALHW HBAS FS 4PRASA 05-5 MERT/ LEH FPRERBERAMES PR BEAT. 57. BAEK 56 HHA, HP HR bd) MLHALHKMHHAE DRESH 05-1 MERT/ LEM FD RFR ERAOES PR RET. 58. RAR 53 HF, LP HR d) CUP R: a) A RA AGA SH, AEA Tat 5 oo OR ES WR MAERHARE, b) 4 Al FACS BLS 3%, Foy AA HR BARE EAC Ba He. 59. RAZR 53 HFK, KPHR)AAES HY LEK AH JE 10'-10° + HE /m1. 60. RAK 53 OHA, KPMG MRHSARADAMER A tin 1 15 EH AR I 61. RALRK 53 HFK, HP MBVR LM, FA FORE ARFRARHRALA, FR a) Kc) AMLELR RAW AGO HE OAK BRR Pa. 62. RAKRK 61 HHH, RPMURASATELE BLA FARREMREEHERERE RAH HAR. 63. RA KK 53 HFK, APH R d) CH MBs HAAS HR Hea RAE IR. 64. LRLE PM HEMLE MA, RARE: a) MBER E-RARRERAH RAWLS Fe BBEEWIES WARE BFA, b) SROEABE SH BT RF A 05 AEH, c) KA ACS Fo tk Ree Hl ES. 65. WEARER, HEFL CUEMLARAMEH RK RAERAR ARG KM Z fa R HK Na Het KE. 66. NLRRP RRC, TRAKCEMLAR AMIE FRR BARE? GBS FROZE RAM RH EME. 367. MEM BARRERA A, PRFAKCEM LARK 84 FY Ae RB AR At BRA Ke Mla Hh thy EAC, 68. NLHBRERG AE, HAFEOCEM LARA DR RB EHBE ERLE WZ ERA Ete. 69. RAZR 68 HGH, LPHRHBRARCRREMB, HE WRRARAM IG, FRPOMSWLESAR. 10. RALRK 68 MFA, LERRHBBLADRBADHBR MH, HRBRLARAMCHEKBRE, FEP-KSWMRAAR B. TL ULERRBR ERG AE, RAAROUM LAR AILEY FRE ERB ERR AMZ ER Hilla By EMG. 12. MRAFRELERRREG EY GK, HURT OM BREACH LFF LEAR BAR FF RE BR BK ZR Tilia Heh Eb. 73. MRAREBEA A A, FRA ORME AR ARIE BRERA REG LMR REBKEZE RAM Hs KA. TA. BALK 65-73 HHH, RPARA RPM LBS, 18. BAER 65-73 HA, HP BRAK He a BP PS Bie. 16. RRA A AIL FH, HSA O-DIAP Frid HMB SLR MIRAE. PASC FoF LUBA F 4B. 77. RRL IH FH, REARAE-O- RRA KA Rie. MEG RR RR FH. 18. RRA DMC FH, AORRLKF HHRRAAARE.Hi A + RA Na i RR ARR KKGGRRREAREAR, WRALERAPRERARP By ANCE hE BEA FR. RA AMR Hele HR (PP) HAE HLH RLAWARAAEMAKARPPAWHEWREH, RA ETP RK, AHETEMAYEHEWRON, Kin, RAR Ae EM LEGA AR AAA BRAT RAH ERM FRB A LEAS SHRM RAR. IR Helle RB HH ATHY YO GR, BPR HEME RAD RORE PHARM RAM AD I. RARE K Hate (AA FP) ME 1926 FRAY, SAP. Perrin VR Bl: SOR AGRARRRADF, WFPFARRPRHZMREAR RHE, MERRHAAD LH $ HEY eK HH (PF. Perrin: “Polarisation de la Iumiere de fluorescence. Vie moyene des molecules dans L’etat exite”, Le Journal de Physique et le Radium, Tome VII, Serie VI, 1926, pgs. 390-401). LE 10 FRE, RADPFHR-HEDANRANREHE BW, RTRABARLRRERG A AH EM LAA RA RHP HMREKHRE KE AAR RK. 1979 , H. Maeda MA TARA FP RAB Keb RKMILZE SK % % 6 MM Mt HR (Maeda HL, PMA, 92, 222-227(1979)), BiG RAARKT Kid DNA fo RNA MEARE (Bolger R. & Checovich W., ZWRA, 17, 585-589, 1994; Yonemura K. & Maeda H., WiG##A, 92, 1297-1303, 1982). FP EG RK RF ABA RS LRA, BRR P, CRE 1EMS WH EKG RRAHEER DER REA SH FEM HA 1S RAAB (Checovich W., Bolger R. & Burke T., X%toR-MBPDTAMEMHIAR, BR, 375, 254-256, 1995). HXMBMLT PPRRMHERRSA, EP 194472 Bl 4id J Mid Wl SW KE RE RR 84) EAC iB KAA TE HH He. Schade, S. 2 #MiET ARRAMALEMAHMAZGBH ve, Ht Rok on th EH 7 HRB (Schade, S. Z., Jolley M., Sarauer B. J., Simonson L. G., it ® Ato REMRRAE YB, UBL, 73, p248-248, 1994). WO93/11249 HG 38 T Ee A yO RF AEG EE 20H, Hf 5 AL HS RB HIP OIG BBR RRA HH. WO98/58085 (AA TA PHEAR AEH 98 #12 A 23 8) a ET MI PARAL HR, (MU de RE) HR RAGA SRR RRR SEP EE BPE) AA. NP ETS RR BPP ERAS, REP RAE TAT A PACS (RAM BAR RLLP RMA MM. ATRERKEBPB, FSERWRA EBT, BRM, CREAR LARA EGA SHE, Ah, BLRRAAT —HRD RD (CFG) MKB, BR TR BGA OB. CLKPRAHLCPARWABKRH BMH BR AT EKEREHSRAM, PRULEREKRRAAANS HRA RE. It HF BRR HEIR DNA Hf Hl Fo BAGG DNA HAE IG KERRASRALHRARRAR AH, HEH RARER FAG A 78 2 So (A oR E RP, MGB eZ) FR OPE ORAL AAA A A. ARP RR PRAT RKGRELMOBRR, KA feHLA E EBS BERAANAEELEAWLS AS BLS EWKEDRHGDRE TH RL RA. UE AIRS RAP BRTRARRRAKA RAE, CLABES ABLSMAEM TH BAP TE Ee RB RAD RA I SN M, ARAM PS RA LA A ER, — AM BEAL SG SEE PR ART HE RK) 0 Bp BUR ZOE TEA. Rt, GRAD FE, (RAPID BO TR HUAPPEKERMES HS, PRSHSR ROR ORH, BH RASNHLLAAPLEGRMHRAARPRE. RARKRAGK FARMAARMEHREBAA RTA PPARALRKS. Kis, RMR RABKRRGH FF HMB, CTRRR Bk, ZR RAHLA, MFRKEPRARBET OLA Phos AGEBSD (BK) HRERE. MRRADF HRA EMER RKDFTRMELG EGARERARKD FRET BKF HOR. MPFRARRM EAA, bo NR a TRE Eh EAD EH RT, AR Ee RET AMS ES 5 Sis TAD Da th Oh AE BP. KRAGE—H GMT SB BALE OE BEA Oa ik, WR Ode: a) KRRAAP REBT RFA FEES b) ARABS 35 Hs AC RB SE TR HR BD c) BSE EH RRR, Fe d) AER ARR A RA RIA. KKARTRERRT EPMO, RRA aERAKYHEMREWERBEEDKEMHRERFA b) HRSA BAES HAE RF A HAE c) DK By th fo d) FEAF EAS RAK FRPP. KRGGKEF GCEAT MAF EMAARABEM LE 3Bt, AAWBRAT ATT AAH RAZ: ERK RRR EH, RFCM ARAB ZS BRIM IH ARAB R Ha EM, BMRREEE, HEF K GMM ATARRBRAM LBS KARI RM R Holi He FMC. EMEP EE, HEFCE ATREPBKAM ZAR KMCHRD, ARM, ARRDK RED RK Heh EA. BRECRREBEL, PRA KCB MAPERSRREB HARAMLHEGRELHSRBHB ERAN EG HE GM PR BAG I OY EIR R HM eK AG. HN A RAE (xyloglucan) AWHRAMB ER, SRF KOI iL APRA RBA HERR BR HAH EA RRS E RBG AR RAG HM ARABI EK RBH Re RK 4b. PWRKRP RBS, HRFKABMIATLRERP SRLS EACR EAR AAR IEA PRR KH Mls He FM. PUM BARRB EH, HRA CMR ATR ERS TIEN ER HARI th PHA FRA OY SE HM HH AL. PME EL, PRG KOM AES RRB WI LARAM IM FU BRB RK Hl Te. PWALFRILERRRRBEH, FHAKOHA ATER FRELBERRBRBEAK GM LAR AMY AFRELERERB WY FE He Mia HE MO, MRAM EL, RG Cie RR eK BRAM FREER RE Eh RAO. ARAL TH ALATA RG Hh BRA EH BH RAW AM, Ait, ARWBAREMILAA a: RAR G H RE FERRARA AK. RAM MLH DRED Fe FERRARA KE. 4RAM MIL LAD Fo FER A HH I. RAB HEIL LBA Fe PEK RH AH ie. RAD AIC RE LER fe FER AA He. RAM BIL BRB PARAM RAK. Fy fk RL BRT HAART R (1) = BRE 85 GE SSE HS (2) =A 0 SE BL; (3) = EERO T fo/K AE; (4)= REISER AL PD BT RHE; O)HLERBERMERK PL, UAE mi RIM RAH. B24AwETA2 HAS. A3tHKBA 3 MAAR, LET MATACRMROHHS (ZA) HS, ARDR(AR) KPBRARBEH RD OLE PK Te Mis Ht BEB. B44 ASKEW 4 MAOER, EKTMATROARER OE 3 (A) aS, AKRAE (B) AMAIA CHAR HUE PRA ia Fe RAR. Ao e5KBWOMKAMAR, LET MATAORSRRER Hite (2), ARSRRRB (1) HAI AAI PRES ER BOL A PR Hs HH te, A6 RS KAA 7 MAMAR, BHT ARR YP BB (PME) RAK AKAGI RIBRLP AA, LG HE (1) oA PME Fo 2mM CaCl, (2) eA PME 42% AeA CaCli, (3) AeA 2mM CaCl, 12% AeA PME Fe (4) Rr PME 4 CaCl, 9 Ht LF AF RRM AP RK He Hy HE de, ATASEBAIMAHAR, PRT MATRAAREBER 4 ZR (ZA) ae, MT EMRE Oy RR (FIR) KPA RAR ARIE RARE AEP RK Hla HG BEB,BPE BML to LL, KLM MAREK He AR BARA FP. ARE FP ARB, ORRAWRR-RF HRAMK, ERAR WAAR FT Oo. RARE ARM LF HH ARH RADE BAR Fo K At HS 1] ii] HD (dw He HE) FY BRM. HDF HE 2 68.5 iP FE OFT) ADF ER TF a ER SPH (41 A), KAT HARM I AAT HX (4 100 WA). BF GREER ELRATRERR, VRTERPUDFARH RAH oR! AF Ab HAAR, BARAGE EL, RE BERR ” BR MA AEA DNA, RNA 4 cDNA WAABA DNA, RNA % cDNA HH Me. EARAWLE LY, RERRRRDRBMA RE RAERHR ALMES RAR HEME DH BR. RRR TR BRB RR. BEARAGE PLP, ABRALA BARBRA ARERR #4 DNA & cDNA Ht BF. EARAGEE LY, RE BEM” BRERA ME, Ree FET KB fo RGA AE BARBY SCR 4A DNA i CDNA HBF. BARRAGE E LP, KR FR Hee Hh gE ow” & REMARK ? CAG BABE DNAX cDNA H BF 64 78 ES. AABRRAHE ERP, RE SE” RRMA KE Zw. BEBRE EKBAREH EEA EP, BERRA AE, to RE a & 4% i (archaeal cell) ALMMBM. THERA LBA k— Pe. Rk MRRP RHEE, wehRF RHE, a ik SK BR GL oh SH, he A, eo 6XS Meribipilus gigantus. BARE FEAR, RFRRKARS MBAR. Jo F LHR, THEMEMEMRELMAS, WIS, KEBAB HAP Sh Pk — HER A DNA SRNA. RE KAO RERAR LS Mish RY RR, ww IRA. BEBKELTALRBERE EY , HAR R oh dW BH 40 fe Sse Gerhardt ¥ (1994) Arse FAG EE Hl Ab we de we, KERR TALK WO97/07205 MHRMUAA LG east RAB Hh ih ta ER. KAKA EEA EP, BERRIES DNA, RNA, cDNA RA LAAAA HS, FAALA APA TMF AA #5), Je KB AA (de Stemmer (1994) & WO 95/17413 APE), MPL % (4e Eisenstadt E 4 (1994) Mik) SAK IA PCR AAS (So Poulsen F (1991) AFR). REBK WETHER, ALARA RBE REA FARO RAH EMULE DHARKR. RKRARTUARMORAP RR. HERIMOR 4 45 R Fo a BH FT AM Ohuchi ¥ (1998) & Ellman $ (1991), & RAKE EAA, WHEARHERRAGRALA. BRL AKR GF, BROTARRERARAY, HEM , RAR AE HGEMBHKAK CEH OM, SAMOS ARERAR CF (Jo F CAPE) HS RRA RHR. 48 ae, BERL GE MBIT KE AKG RAL ERR Fi Wy 5 AE 4 Sa He. REGGE MBALRERRRERBR FR PP pik ik EBS RAS (ABR EM RARLERAREWLS 7MM). BMORE TAR MMAR, AAihitd Christiansen ¥ (1997) & Stoss $ (1997) HERA HEATER A RRA. ERB ADEE KARP, 8 ECR ih KI aie. 9b, Bk 49 WHER ElectroMAX DHLOB(GibcoBRL/Life technologies, UK) 49 18 % X Ji +f i # #49, to Diderichsen ¥ (1990) PFI a4 SJ2 AMAR. HERA REMMTARF BH RR HK, wDHEF RH RAF RA RH. HRRHLBMEAMA, wih wae. HERB SHE TRA CHFEWERALE. Sm AE BR IER DNA HAR LH OY RIE TH de Pitcher # (1989), Dretzen, G ¥ (1981), WO 94/19454, 3969, Diderichsen ¥ (1990). WKS HSRAFRRRASRALAH TABS Stemmer (1994) % WO 95/17413. HRA MA HE 33 HAA RB XA DNA CDNA HBF RE 2g FRAGA B40, BIL Sambrook F, 1989, Ausubel 4, 1995 4# Harwood 4 Cutting, 1990. BARGRAHEBEAR YP, RARER KBBSA BR HATH HRM GARAGAN, HPRERAAREG RFRA BRD RARRIL). KAORBAHTHRALE, OHKE, FREE, AF HER, SEEK zeocin. BAKU GA —-PRBE EARP, TRA W094/19454, 3969 Fo Diderichsen 4 (1990) Afi& 0 KF HHH He 5 pSJ1678 AH DNA He 4M SJ2 KM BE. RA, TKI AH pZEro-2 (Invitrogen, CA, USA).WELMHE, ALRGRKT HRRGADLE RE REA Oh Tik, RRA Bie: a) ARE ARP REBT BFA UP EAM, D) RABIES HH KEWILE WR BRK REBAR AR BL, c) MRR AW RAR, Fe d) FF R Ae RA A HAG SMR AIK AK, SRR OAK RARPRE RR (Go LX), HR FRR BAS: a) WRERA RAGA CH, b) HXEMRAKGPHAAAA MSE. c) Fe DH KH Be, d) IMB R/ HAE SH KH RIK A Leth. )ESRRLES PY HEMKRSMARAER DY RA MRE, LHEBRS DH RAWR-RRB, 8) MEK AWE WA BER RAMA PP EU, RAROORARE, PERERAAKHBRRE, MM, Bh, BA, BRERAF TES ATT HA 1 AEH ag (C1) +(2)+@)=HSRALE, A)=9H, PH FHR/ MB, (5)=5 RIM RAR BIA FP). BUH AWLOW RE REHAB A DO EAR PP wR ATRL, GE HHH PP PRTRMRAEG. Ht, RAASAH AGE tg PT Hy RH Ba HR BLAS Ya ACS HEH ER Bl 04 EEPASARA PP LM (Hp FL PHALA MSP”). RABAT HEMEERR SARAH FPR BR, REE ARH PEASE A, RG RHR EB ERR EE, PRABAGLY, PTD RLAPORA HB. wit FMM PCR 9BARU BKIME RE R/ BEG HORA GP THD BORA 1 (IR Ohuchi ¥ (1998) p4340 & Ellman (1991), HAAAHA SA). HATH Re), ff ghMKEKREHATLMBAKARH 4 FBR. SARAGRREBEA RA MRK RR Go ELAR) OY, Hse A EMRE: a) BG Hi eA S AAT RF A 04 oH, b) HRP FRARAMEE , c) Ff ADH 6 Ba HAS do BA Ah SE BP AES HY ESE, DESARLES PS RH ME, eo) Ha 4 RAW MR —- RRB, f) Hit 5 MORK LBKOWR VER KEW IAH FP RA. RAWDMORERE, PERFALAALMHBRRE, Wi, WA, BK, PRBRAFTE SMTA 1 AHH R af ((1) + (2) + (3) =F Bi FoR; (4)=9 Bs (5)=5 RAW M-RRBIF #eR1 FP). BRB H PRA RP, FNC BG Hi Fo HARI RAR KRY MOREETRUSHRELAAD BMRA FDRA. BH SARA P ES 05 nF) HK A 0. 3-10, KKH 0. 3-5, $e 0. 3-1. AEM IR ASE RAR YP BEAT 0 BO LP a HD) HR LEK AT 225. CHHRABREHA 10-60T, KikA 30-50C, 4 37C, Me pH MH 4-10 Zi], HHA 6-8. KR BHM ARAPLRRHSRKT HPRLEREHER, RRR A 15-60 4h, te 40 4p. SREARARLMREHRM, RPMRKTOAR ARERR RHRALEM, GTUPAMARART ROSA E wR GARE PORERMLOREK, RELMOTEFAPOREER AR, Ah, PLAAATALAZADHRLEMLEARARE MF 6 EA HEA EEE EAT AR BT BO BB BIE HH. 0GUA ASE RAT ATA (BL PH) RA BERERAATHEOE RP. MUHA LAHLARHEPRARBERAMES, eR MAL PTD ke (AL PRS). KA, FH OS HL Be WO98/58085 (F] ARKH AKA) RAR JA FACS SRL 4T AM, Alb, FR b) GI FH HR: 1) AG RE HEB AGA SH, ap So a HG A eH 5 Se A RE BE KER ABR, 2) AJA FACS (U6 3%, AR FO HARA RR EK AG Sa. GRP AAMRABMSAES PH LEKA, RRS 10-10 TE /m1. WREAG RMB, THRLRHA EER? AT Hx RRALH, —M BRR M 50-100 RAA 96 LH FFU. ARIA EF — MERE APRA EH SE a, RAE EP BORLER ELH RMBLE, BTRAALKRRANS HABRA RY BL OY Sip 3 Se HL 04 TEE LA-PRANEKARY, CHHRRRREHA 10-60T, Kk A 30-50C, Jo 37C, mH pH HAA 4-10 Zi], HAA 6-8. HK BS HLS Re LED ERERAMRAR HER. SRERRA BERBER, LEHR T SAE ARERR RHRALEPRAERMREM, TESRERERAR MAAR BEBRM IRR. BH HORA PUMREK PARA H/T RESHESHARRHLEHSORH, AHALFRPRBAK EH LE. RH MAA 40-90 bat, do 48 ba, AAR De FAA Yo FE E50 RD Fo dh HM EH. SRELRASHRALHHREMREHR, EPH BE ME POSKRARMLM, AARUGHRRLETEY, BEAR ERR ARE BE MBMAKMRFRPUARR, HH Fo / KG A. RM AIS PA st ARO HE a SAA nRE HAF AK AMA EAH AE RS) bw. MUM REG MEM FORA EET MS PP OME we (1 PF v9 A) P, Se Bibby Sterilin AMAA, UK &K Dynex Technologies, VA, USA §#2GMMw. Wit He ABAD } wrAw TRA Hh Hin BH / HE — KS AIL AL PP AE A Re RE AR RAR LETH ERD ADE W EE (Wi TS RY RRM BBR HAIL RR). EMRE SE EK MZ 1 ER FERAL MOPBHKLY, HEKAAA TRAAAM PPAR. H EMS RR EAS, TAM ALAR BRE WAGE DNS HE ERR P. He, BHRP RAW AH REWREURADLSH ERA WRB. BES AWMKEMRE MRED REMAK DH RACH PP wR ATR, MUH HH PRRTHMRHRK. HH, RAASHH RERBETHRAW KREG EMIS HRS BFA 0 PTS AE PP RK Is Fo FL PHMRLA * MP”). TERAFEMREMPRAMRZMKRER OEE TE RAT BR. ARKH EEA EP, M pH PERHFRRAD KR FEBS LAHR ERE. RBETAF ROBE HM EH FAT oR 4&5 BBE (So 30°C AF K 20 AF) Ky M (So 60°C ALEK 70T WL), 46 pH de 5 AF 4 AF) AH pH Ge 9 LER 10UE), KABHAF BR, RERSMwETNHAA), RRATERM MH EWES. ite, weRAFRHEMULEMLZARBZENLOH, PTARRE HRB, GHRHHREHARAERRT RARELY EW EOH ARH RAWARB. RAG MAG PEA FA PP RAG GY HH / TERA RIK HR, AFRLEEAFRRRAWAK PP HEMFZ, LUMLG $ Ht PMEKFT 3.5. RERFAEAARH HED, CHT RBRED LPHASwWEFH—, GULLS LAMM ZHE FD (REA 27 2fo 28). SRI TEI RAIL Jo TEM) ME, AN 96 ZL Bite te ERA 2 E3K. Ri, —H RRP HRB (dw KA AF IC GER 4Fe PASC) PPA TRAMEH, RAS EB THAR Ei Z. WT RMAR AMR TH RP Ah RS, ek RE AERA, PRAM MEME TREK FRA EAM, BI SBE. BSG, 5 9 5 Sn ERR AR OY DNA SAE ER RE ARP REARPRERBARKREREMBAREARPRAEGRALH, KP RARAHAREDH RADAR EAE AER al. ERB —D BRR EEA EP, PAHO MET PPR IH WRAL P PP RKP TAHEM EARS AWKSM A th BI. BABAR DR KARP, Mes RAK AHH HRM, RATA AH MZ SE PP RUREM RARE BLEW BRE (ARM). HAOREPHRA PP RMKRKPHAHAREW ML, Hie Jolley Consulting and Research #4, IL, USA. HERS LAMA R Checovich ¥ (1995). SEAMS PH, BUA APR ARALRELARGH FETAEERRG EMER GBA SoH R.A S SRA P a Eat ALT SRA RE Rp 4a Be oT ik 5] EE 6. EDRE KRREFLP HEME TARRERERRPREFLA LULEAAHEMRSH. BARAK TEARY, EULSCHTRREAR, FRR Ak.Bik EWE RA, HDR EE AOD FA ERE BPLERA HAA (1992), Academic RADA, 1992, THE AFAIK, to HACE RB (EC 1.-.-.-), HBR (EC 2.-.-.), K WR (EC 3.-.-.-), ROBE(EC 4.-.-.-), JPAB(EC 5.-.-.-), 4&4 (EC 6. -. -. -) PERM TAR Aa, sa~, RR-KR RRA, Ay Rp WHR, A-SI, BUBAMS. Kt, EBKEBHALAMRERABFRR, WRERRE EAR, SRRK, CMAALAWH TARA. KERRY EWCSW A BPH ICS, Io Oba, PPh ae DRAB Riek SMEAR KHSH. EXERT, £Mtt SW LR RMP HEH, BA SRSA Ee PR AG. Bio, WHER OH —-EBRAEE YH AARBARS MR, iio REG MA, PSS RKAMKAORERA ER. BM eae Pe, REA eS, ARR RMR, ER 4§ 4 VE (Simonen fo Palva(1993)). — 2 XE BAB Ae Me EI PURLERR EMG, DG TRAE (Gotz F (1998) # Pugsley # (1990)). RH IR KERLFLPOHRAMR TAL AL RARAR ERR A RY ETAMAEWLEH, AARAGE ERP, RR RAR TASHTEHRAMRPRTAH, BAUFLRT MRE RRABRLASH PP, PRRERGRH AGRA. MFR FRLRAWAERLEMHF EGS, RFLAALABA: DEBE RAM KY FRAY (RAM) LM ALK H, HA KEKERAAOHAERARRHAHER, Bb, LARWOHLE XP, ANGRLW KMART BRHG TAS EWKLSCUR BR LER (LHRAWKAESARADPERAD, CNL I 4Lb A — SE). Li) MEA AH RRARATRH ES ORAWAHAS REM, KRAMKHARKNGLEG LL EAP TS EWKEWR RAR RA. Be, TRAM RRR RH do He fp RU HE POF i oo Fo Ae Sn 18 SRA RA. iii) RAMKRSH ABE HREM LL SAP THEMRKEDR EH RZ Hy AAR RA RRA RS A, PEAT SPRDHARAMLON, iv) MAU TRTMRAHABRGA RGA, PP RMR FHA HAMS, PREM AT RRERY. V)FPEMAKRAEDHAIRM A, PRAHAZAAAD SH WwAR EK EARP, RAM MRRRR AMI (KAM) HIT AMDT. AMIFTARSRK, ERRAER, HTR-FR— RUEMRAREW HL. BMA FLARE TRBS EBRD E 0. 01-10% 7 HAE SH ES (ARIZFAE IL). MIE A 0.5-5%, te 2. RACH TRLR HR, SFRAARSH AMP FAS. AMPFRAAZGK, HK, REG, BEG, HH, SRAR , PM LRARER. AMP TET RAMBARALE PR 5 EE RS RRA, ARDW SM, HERSEK. LRAT TETATPRESRKHAMF. KAHZOREBEA, BEANE, WR, GEA, KZ EG, SLEEK, $LERK, ALRKAAWEK, wPAREA DK/97/0879 & W099/03498 PRB ay BRAT. ik AP he. KEG SRTALGRBE, PRP ARR er ARR /ALR SP), WH ARM), RR/SRERBER, FHKE (CO Schulein (1997) P38 #5 Fl SERIA RA) AMAT AMAA RE. KAHFHRERABRHE, ARB, MEMDRBAMBARR. HERA (ELE SRR) PRELERR Go AFRFELBERR 1sHe) BAA. KEAAHAMAFRMRI FT, wT AAKERR RG BRED EF Hh. BMAF THMECRRBRER. AETHER, A, BH KPA MRR, ET MG AGF A ALE A Fo / HAE. 451d. H. 350-700nm HEA A, te 400-600nm XA RK HL WARE, RAR KA 36 HEA 450-800nm, 4» 500-700nm. KAI Ss AWD FT REX WRI S AMD TREE RAIL Z ie] HE BRA A 20-50 HK, KHMER. Hi Fo MA FZ] HHA A MSE, SMH ICH AH eR H, MRZAMAF OBA, Aithipith BALES ERE RAMA FEA (HARMS F th RAH) HEH. MIB RARRH A 1-7 WY, HRA 3-5 WH. Bik 4 HID BODIPY HEM, RAKRKMAM, FHRRABRHK GAM, FRAREGE (SoHRBP RR), SOPRA OS THA Molecular Probes, OR, USA) AR 7% #X#H( TA Amershan, Buckinghamshire, UK). RAR (BMAF fo RABI HRS) BAA Hh ee RALIE A 50-500mP (& 4 3h #4), 3A 100-300mP, de 200mP. KRPECEHP LEADER MAN GHHRAD MR. AR 3809 RAW AA RA B46 I AE, Bl to HA ERE HK OS Af HEE (PASC) KAP RAE (CMC) Tak de 6-DTAF 6-RAK-KRSR) HRARH . FARAAREMA KAGE: DFFREAAMAWRFARES FEKRAREPR Ps i) REFRE MG FREAK RA AR B/A; 111) MAAR RPM PD HBC FRER RATED HARA HRA iv) PHRAMIEHARERE TED. A-TRAGRAMKAARAMMZARERLBRR. Hiv, RERELERRRB T Ko 6-DIAR(6-RAK-~H=AR) HRAA Ai, PERARFRERLBRRRENA KG: IERERER 6BERR RS fo /REMTRASKPRM PS 1) RAFAH RK PRT LMR BRS RAM RB/KM; iii) PP RAR RADA; iv) DP FARRAR PD HARI REBT LBM RH RB 2H LFRFELEERRE. A-VKAGRAWRAARAM BLOWS RR. to, #4 WEA T HK to 6-DTAP (6-RAKA-RAB) HKAM MIA. PERE FHL SERA HK AG: 1) EF GUN ERA RF fe / RBM FBS AGP Ps Li) RAE RG ELH ERS RAR B/ BM; iii) BEBE P DB AR TIL BR HR IS EH SRA. RPK RAWAM RA RAM MICH ARAB. Hoe, KR Vie BARI ARAAW RAMI. PERAKRBHTEE 46: I MKB GS fo /KERTRESRKP RPS 11) RARER 4G; 111) RAR ARBS RAM RB/KRB; iv) DH HBR Rte BARI ARAB. FAB RA MR RAD AIH AR RA. de, KA Rite T to RAKES RAAB RAM. PERAK RBAA KG: DHAKBRBR I He /RERTSRARM OKADA PS ii) BB / ACI E098 DP FC OK A RAB LAs 11i) RAH AR RBS RAMRB/RE; iv) PRP RIAwIEG KA RM. FDRG RAW MRRA RAM BIL MARR. Bie, FAA RAT tho RH E-S- REAM RAD. FERAMBAR MM H&G: 1) HM BARR FS fe / ABATE OH RESP BPS ii) PB / RACH EBD AC Ms RAD; iii) RA MFA RS RAM RB/RE; iv) PRP AB ILE RA. APR RAM HLM KH DAI RR/ SRF LBBB. tio, TR Ade 1-OR-3- (= PRRK AK) RH SMe RH wy PE WH, Ae RAR-5- RRA RA DPEREHRRAD. FER 7ARBAF EOE: iI) HRKRAF/ARERTSRPM PS 1) RR KEG RA Fo Pf PH B/ RB iv) DH PIR BACB IC RB, LEME ERA FP RETATARA ERA HADES HZOEK EP PRE R BEE . PA Fe AR FE ARE AK BRAGA AT SAH KE RR AR 4, PHA F 5) He 2 HR RM A Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH: 1999-02-03; #284 RCS 242B; DSM RFA 12663, 1999-02-03; #44 RCS 245B; DSM RRA 12664, 1999-02-15; 453% # 404F HW MB208; DSM RRMA 12682, 1999-02-26; #7047 4s DSM RRA A 12708, 1999-02-26; #ieA MMH; DSM RMF A 12709. AER BL ALAR LBS ARR PH LAP HEI DNA TEE oR. Kone, whit Pitcher F, 1989 MAM AD BRAM DNA. HE FHA 25 Fo 26 P, APRESS SauSA 2 14 LK A 4a DNA, Wt A 0. THB RBRRE BREAK DAD H. 3st DEAR #REBK(Dretzen, GF, 1981) PRADA 2 FE 7kb HH BF. 48% DNA HERE BamHI] HUH pSJ1678(R WO 94/19454, 3969) M# DNA, RABRRS WHAM BH SJ2(Diderichsen ¥ (1990)). Me KA SJ2 fale, RT MEMS, (AM A BIO-RAD % Gene Pulser™M % FARM w FILM ATH. B Sambrook F, 181989, Ausubel ¥, 1995 4 Harwood 4 Cutting, 1990 Mie RTH A HAF EnELH. 4 XH.) 27-29 , BFR pZErO0-2 (Invitrogen, CA, USA) He CA: ARE Sousa SHA EAR DNA, BG Ht RR WREAT ADP ADH. 4G 2 E Skb HH HAREM Bamll HL % plEr0-2 P. MURWERHAHFTERTRFL, MERRSW 4£4 ElectroMAX DH10B (GibcoBRL/Life technologies, UK). #4 RAR LB SHA PACH IE HR OO DH, APM EREA 15%, HEPAT. GAH TSA 10g /ml FARE HHS LB AR FREAM RAD ED AP OECTA. FDA BH A RBH PE TK AF SJ2 ATRAEFR, RARMA S20 HR (stacker) M Titertek Microdrop ## (Life Technologies, MD, USA) 4 43b3F 4G XE KALP. ATHLEKS HBB ERMA, KAT Platemate 4 HH RE (Matrix Technologies, MA, USA) RH. # LB A 3A (5g/1 NaCl, 5g/1 HAIR (Difco, MI, USA), 10g/1 RM GK (Difco, MI, USA)) PHRMA MAB SJ2 HFAK H. 437, #Nunclon MicroWell 96F FAK (NUNC, J+) LHe HK Ma BCT REA, HMA 200rpm MRR 3A, RGRAT HK. 4 PPM LM, MUL 50pl HKFARKES 150p1l CHRAW BERR REG FTARLLES RAKES. WER Klee ATHMASEPHRARR, RATAKERERGERH LS50B (Perkin Elmer, CT, USA), ATHRM AMA PH RAR, KH FLT FPM-2(Jolley Consulting and Research 28], IL, USA). ¥ AA ALA LAA POA STE LR OH ZRRA EEA WE 4%, RAT BH KD RMB: Sterilin + RIM (Bibby Sterilin 19AFR], UK) 4 Microfluor U-s& 4 (Dynex Technologies, VA, USA). ATEMMABAPORLAA, BAMKREAS, He 485nm HOT Vt, FILA 22nm, MARHBASB, 1 520nm HHT Ut, F 344 30nm. ATEMRARPHIAR, BAMKRBASB, KH 546m HET Vit, PIMA 12nm, ABGARHBAS, HK 590nm HHA iit, PIA 35nm. TRAE BRT Mt EAM AK. EMR EMLABREM HT AGRERATAABARE, je 4H RH LS5OB (Perkin-Elmer 24], CT, USA), RT MA MMH SAX, Je Polarstar (BMG, &H)) ARAM FR ARH FACS (AX WCi6 Fa HED HKAR) (Arndt D. J ¥ (1976) 4 Lindmo, T 4H. B. Steen, 1977). REP AEA Behe AAR, PPR AW, BODIPYO-TR Fo tk & HM Scie Ay PE 34% A Molecular Probes, OR, USA. wf 45 ia a5 % BEARAG Ho EAPBRARA ICH tS RMA A Sigma, MO, USA, AE1h42 RIK (3%) #5 44% KAKA A Copenhagen Pectin, Lille Skensved, #rH. MAL A 17. 4th = RRB A Herbstreith KG, @H. ARM Ka AEA AREA A Sigma, MO, USA. 7 A SAR OM Ho 3b Fo 7 A tamaring HF 4 AKA RA (RMAF) AH A Megazyme (KK, WAAL). FALZH ARH (111 B) HHA RKRH LRRD T 4 4 Sigma, MO, USA. 48 Locus 2K + 4LHS RAE TH A Sigma, MO, USA. 48+ SLSR MARAE TF A Copenhagen Pectin, Lille Skensved, Jr. FH Schulein (1997) ApH A RBM oF #. Wit Tada F (1989) MR, AA DAW SO ROAM BARD Re aSERAT FAM, CAR WO9414952 PHM ELEC RS I, IL, TLL; +e W094/ 21786 ETH HRW EH RRRSB, to ALP HT DK/97/1344 FEE TA A He FO RRR SB; do WO94/20611 Mit Tt 8 HO wh HE os M41 RRA; Jw WO94/14953 BTA Rew EH KR (A RES IL); THA Novo Nordisk A/S, #4) #8 (Termamy1®); 4 Christgau ¥, 1996 PRT FF BATE th EO RBC PG; dor WO9G /23366 PERTH A ERE ( Phytophthora cactorum) H#BRMBRE; Je Pry F, 1992 Hk RAG YE BARD Heh + ERA REA ER, TFA Novo Nordisk A/S, Jt & ARAM (BioFeed® Wheat); 4 W094/25576 PRR TH BR OBR, FFA Novo Nordisk A/S 4H Novamyl; # Shen ¥ (1988) Mt ié F443] 4 C. thermosulfurogenes B- BEAD AE; te WO94/ 20612 PRET FE HRI a HE OS AEE ELE RR BM; to W096/29397 AFR A Humicola insolens #4 Cel45 ##E Me. BP-X 38-3p 3K He F RS BP-X 3S HA: HH 100g ZH, 50g KH Fo 800mg BAN 32.9} (T4FF A Novo Nordisk A/S, ArH) RSF 800m] KP, Are REM 10C, Ri 30 PH. AAAHLAP mA 10g REABA (MD-Food, fr). SA RERAM MA 20g 2H, 9g BRA (MERCK), 0.1g pluronic PE 6100(BASF) AAR 1000m1 *. TY EHR #& Ausubel ¥ (1995) Mk #1 & TY 28H. Ke 33 F FER RHE AM BARR: KAA 1: IEA A RE He Pry ¥ (1997) FR ARRI AWA EK A RAB. KA 2: LAF RC IK A RAE 2