PEIIEH3tr`
Transcription
PEIIEH3tr`
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Ceprt or IBcneABaHrfl B ToBa HanpaBJrenrenoxa3Bar, qe pacrrrer'rHure crcreMfi 3a eKcnpeclc qe npeanaraTpeauqa npeanMcTBanpea KrracnqecKrTrE, ca nphBneKaTeJrEx nopaax o6cTorTencTBoTo, u3non3Bar{n ApoxAh, 6aKTepriarH! r xrBorrrHcKtt rorerK}r, Te oc}rryp.raar no-rorrMa 6e3onacHocr 3a nPor,r3BoacTBo Ha (bapMaqeBTr,rltHr 6eJ1mqx, nopaA! ibaKra qe He clAbpxar qoBe Rrr ldu xr{BoTl.rscxrj naroreHu, oHkoreHfirj AHK nocneAosarenHocru fi eruloroKcHHri, xohro 6uxa Mormr Ia 3aMapctr KpatHrt npoAytT, Kar_roH Bb3MoxHocT3a noryrraBaHeHa aBT€HTuqeH npoAyKr, Ee,'rrbqHam cnlfre3a e KoHcepBarr,rBeH npouec npfi Bucurhre eoKapr,rorH,mKa qe pacrr4TerHme rorefir] Mor-aT aa ocriryprT npaBluHa c1pyl(rypHa TpeT[LrHa ri qeTBapTuqrJa opraHrsaqur Ha peKoM6rHaHTHwreqoBeuKri 6enft,uH, H€o6xoAr.rMr, 3a rixtara 6Honor[qHa aKruBHocr /3a pa3 jKa or 6arreprlaanure KnerKlr, xlaero 6srrr,urne xe ycnrBar aa ce HarBHar npzrBl,rJlioB npocrpaHcrBoro H BnocneacTsqeaeapalnpaT unr ce arq/MynnpaTB Hepa3TBoplrMri renqa Ha BxrrroqBaHe/, flopaar pa3Hoo6pa3HureoyHKul.ru, Korro naKTo(DeprHrr /HapuqaH oule naKrorpaFcQeprlH LI npnHaalexaul K6M fpynata xerrr3o-cslp3Baun DtHl{orlpoTerHri/ npuTexaBa, HeroBaTaxpaHrrrerHa H OapMaqeauvxa croixocr npe: nocreaHrffe roar,rH, ce Ha6JtoAaBaBce rro-3acr,rrBalqce rHTepec KEM pa3paboTBaHeTo Ha HoBu e(cnpecroHHr{crcTeMr,r,korro MoraT aa oc[ryprT roJreMHKo,'rl,tqecTBa qoBerxKfinaKro(bepHH B 6riorora.rso aKruBeHpeKoM6uHaHTeH e cuHTe3rjpaH 6errr,K. PeKoM6rfiaHT€H 1993/, Pichia pastoris /Jiang et al, 2008/ r B rb6lr Sachcaromices cerevisiae /Liang et al, apoxnr Aspergillus orizae. B;npeKu ycnerxuoro npou3BoacrBoHa peKoM6unaareanarrorlepnx B Aspergillus, Toii ce F)d(qae or npoabJDrorerHonpeqfcraaHe,KoeroocKbnrsanpo,qyma(aro xpaH[TerHaao6aBKa, flpor.r3BoacrBoro Ha pekoM6x8arrrH! 6enfl,ur.r s xksorrHcr(rj cucreMr., H3l,rcKBanpoAb?'DKxrerHt r,r cKrnocTpyBarqfiTexHororrx 3a noly{aBaHe fi npeq[cTBaHeHa KpaiH r npoA),,KrH e Hantl]e BeporTHocT 3a 3aMrpcrBaHe c Bhpycul, oHKoreHHu AHK nocneaoBaTenHocTx, qoBeurKri naroo(DeprH ycnerxHo e noJryqeHB pa3jrHqHrpacrlrrerlHH BlrAoBe- TrorIoH /Salmon et al., 1998/, Kapro$ /Chong and Langidge, 2000/, loruar /Anzai et al., 2000/, opw l{trzai et al., 2000; Takase et al.,2005/,qapeBrrra /Samyn-Petit et aI.,2001/, xeHueH,4(wonet al.,2003/,r66n|(a/Schneider et al.,2006/. CpaBHrrerHr.rrr aHajrr3 Ha npeahMcrBara fi Heaocrarulure Ha !3non3BaHrre pacrmerHt cucTeMu3a npoayrlllpaHeHa pe(oM6uHameBqoBeuKt nal(To4epuHnoKa3s4rrenoaxoaflq Kali.qnaaT 3a qe pa3pa6orBaHeroHa HoBa li €(beKTrBHa mKaBa e nrcqepHamMedikago sativa L,/. floKa3aHoe, TpaHcfeHH! pacTeHrr nloqepHa, 3a pa3nxKa oT ocTaHarhTe pacTrTeJrH, cucTeMu rcHeprpaT oTHoorTeJrHo enHorunHr.,N- nr'ruxaHoBH cTpyrcTt?r,r /Bardor et al., 2003/. 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Or rpaHcoopMrpaHnre169 excnnadTa/45 or nucr H 124 or ntcrHa coMarH.reH eM6prorcH€ttc ca pereHepupaHu84 pacreliE , or Kotiro caMo6 AprxKa-/qpe3 uHAupeKTeH I,IM c npor.{3xoatrrcTHa ,qpa'tka ca BKopeHeHrj Ha ceJreKTuBHa cpeaa - noKa3aTen3a TpaHcreHHaTa ekcluaHTu or rlloqepHa ca npupoaa. 3a noa6op Ha TpaHcreHHrfTe T6raHu, TpaHcOopMrpaHUTe no,uno)neHuHa npeKbcHaraaByeranHaceneKeqrr c L-iFoc(bHHorPuuHH:I eran c LTnrriBHpaHeHa Ha pacreHwrraHa ceJreKTHBHa cJreanpoBe)KAaHe B5ll cpeaa3a KajrycoreH$nc/2m8/l/ HenocpeAcrBeflo Tpa8c(bopMaqurm r npoArr'rxHTejrHocT ga ceJIerTuBHI{t HarI,tcK Ao notBaTa Ha luoep€HrlrpaHH ruro6yrapHr.r cr).KrypH. Pa3BHTuero, 3peeHEro rl npeBpr'{aHero Ha coMarHqHure eM6pldotlata B pefeHepaHTnce ocbuecTByBaB oTcl'cTBteHa ceIeKTlrBeH aneHT,ll eTan Ha cejlexqltt e npoBeaeHHa hr.rBoBKoperurBaHe Ha QopMupaHfiTepacTeHur,flpeKbcBaHeTo Ha cejrexurrrTalaBa Br3MoxHocTHa TpaHcreHHrne TlKaHlr aa ce B!3cTaHoBrT fi oT Trx aa pereHepupaT ,ao6pe o6oco6eHlj pacTenlir. ycraHoBeHara B Te3u eKcneprMeHTr{TorcsqHocr Ha L-tloct|orpuqfiHa B cpaBHurelruo rlrcKa roHuerrrpaql-rt /2 mg/l/ 3a HerpaHcoopMhpaHne Tr,KaHur e(beKrriBHrTeKoHqempaqhn 3a ceJret(qrt Ha rpaHcfeHHr pacTeHat/ Mexay 0,5 x 3 mg/l/ KopenHpaT c aaHHuTeHa apyt-Hll3cneaoBaTenH /l\4a et al., 2011tHM et al..2012/. B peynrar or reHerrlrHa rpaHcoopMaqfir nocp€AcrBoM Agrobactedum tumefaciens x pacrurenHa pereHepaqru qpe3 nHAxpeKrEHcoMarrrreH eM6puorcHe3hcca noJrJAeH, 5 K'roHa TpaHcrcHHapacreHl,trr'r6qepHacopr O6HoBa l0 B rcHoMa Ha KoFro e AoKa3aHonpucbcrBrero Ha qoBeurKa naKro(bepr.rHoeax{HK. ycmHoBeHa e rpaHc(bopMaqxoHHaeoekrsBHocr or noprArra |]a 4,03% /lrpa3ela B npoqeHTexcfirraHTu, or kouToHa cerexriBHa cpela ca pereHeprpanr eM6phoreutu qe kanycu/. IIopaAx o6crorrenHocrBoro, ca Hanxqe rry6n[Kaqut 3a noJr)AaBaHeHa peKoM6tHar{THr,t 6earrUr.r s pacrenrir lroqepHa, B xorTo ne e oTrreTeHa TpaHc(DopMaquoHHaTa e(DeKruBHocT /Bardor et al., 2003; Dadmehr et a1.,2011/ noryqeHaTaB Hacrortqrr AucepraulioHeHTpyA etDeKrxBHocrHe e clnocmBrMa BaB Bcriqr(r cnyqar H BlnpeKu HHcKam i crofiHocr p8yrrarHTe or npoBeAeHara reHerr.rqHa rpaHcQopMaqurHa eM6pHoreHHu pacrrulr.trluHl,rllnrcuepHac K,&{K 3a n HeopraHu3upaHo qoBeuKfiraKro(beprHca opurrHanHh,Tr,fi KaToToBae nlpBijr onnr 3a rrnor3BaHeHaJlroqepHamKaTo '{oserxKhnaKrooeprH. KorxqecrBoro Ha eKcnpecrioHHa cHcreMa3a rlpo,DJnlxpaHe Ha peKoM6uHaHTeH peKoM6uHaHrHxr nporeaH B pacrrrenHxre eKcrpaKru HarpaHcreHHurepac,Ie4:,te 32,85P9A u 47,15 g/g o6q 6ernx *orserHo 3a KnoHoBe 35 u 53. J Pa3nxgHri npoyqBaHxr noKa3Bar, qe aKTuBHocrm u ekcnpecrutTa no,q Korrrpona Ha KOHCTLITyT'BHATe npOMOTOpCepa3nfi.IaAa3HaqI{re.nHO B 3aBI,tCllMOCT OTpaCTUTenHHJI BI,IA,OpraHIlTer.t (Da3araHa pa3Bnrrero Ha pacreHurra. Pa3JruqduhfiBa Ha eKcnpecur ga pe(oM6nHaHTeH qoBeuKu naKroQepxH B Jlr.rcTHaTbKaH noA KorrTpoJraHa KoHcTrryT[BHr.rr 34SCaMV npouorop ca oT'reTeHr rl npr.rApyIH pacrHrerHrr BuAoBerrorroH /0,1-0,8% or o6unr pa3tBopl.tM6enGR /Li et al.,20041 tr (aproo /0,01 - 0,lo/o lchong and Langridge, 2000/. flpr Haco.reHaeKcnpecfir Ha peKoM6!fiarffiln 6ertr,x n ceuena or opr3, qpe3 r,r3rrortBaHeHarLkaHHo cneur{brlqHr npoMoropu ca nocrurHar}t Br,rcoKlr HuBaHa€KcnpectulAo 25% or o6urar pa:raoprM 6o'rTbK/Huang et al., 2004/. Teslr excneplwexra"rxxpe3yrrarl Tpr6Baaa ce orqnrar nph 6rAeuryrercnepaueuu sa onTuMr3hpaHe HnBam Ha eKcnpecl,t npl,l u3not3BaHe Ha ttoqepHaTa KaTo HoBa pacTrrenHa eKcnpecuoHHacl,lcTeMa. IorBrpAeHo e npucbcrBriero B JrHcrHH excrpaxru or nroqepHa qpeg wastem blot Ha peKoM6HHaHTeH 6eJrrbkc MonexynHaMaca80kDa, Korro c6mBercTBada npeqtcreHur .,raKro(bepuH m qoaeuko Mrnl(o. 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