Paper Científico-Transformação Genética em Escherichia coli

Wdjitðrfn kd [rigijan Urétf`n ‐ Jignritðrfn Igdrtn kn FUI[FMTU, Unrtn

[rihslnrmiãàn cdhætf`i dm Ds`adr`afi @njf Fhsdrãàn d dxprdssàn kn cdhd CLU itrivæs kn vd`tnr pCJN

@jéukfi Lrih`n | @jéukfi Jngn | Dkcir @nutn | Pnlfi Nsðrfn

Wdsumn Ipðs `nh`juíki i I`tfvfkikd Urétf`i Jignritnrfij Gi`tærfis Ljunrds`dhtds, hn jignritðrfn igdrtn kn FUI[FMTU, djignrnu-sd n sdcufhtd rdjitðrfn, hn quij sd ds`jird`dm ns `nh`dftns kd [æ`hf`i kn KHI rd`nmgfhihtd d kd `nmn dsti pdrmftd fhkuzfr hnvis `iri`tdrístf`is ldhntípf`is dm gi`tærfis Ds`adrf`afi @njf , utfjfzihkn `nmn vd`tnr, cdhdtf`imdhtd mnkflf`ikn itrivæs kd dhzfmis kd rdstrfãàn, n pjismíkdn pCJN2 d n cdhd rdspnhsévdj pdji ljunrds`ïh`fi CLU (Crddh Ljunrds`dht Urntdfh). Ds`jird`d ifhki n `nh`dftn kd [æ`hf`i kd Crim, Crim hdcitfvn d Crim pnsftfvn d ifhki is vihticdhs kd tdr `nhad`fmdhtn ki `nmpnsfãàn ki mdmgrihi gi`tdrfihi.

Igstri`t Nh`d `nmpjdtdk tad Uri`tf`ij I`tfvfty Ljunrds`dht Gi`tdrfi, fh tad npdh jignritnry nl FUI[FMTU, tafs rdpnrt wis wrfttdh tn `jdir `nh`dpts su`a is rd`nmgfhiht KHI td`ahfqud ihk anw ft miy fhku`d hdw padhntypf` `airi`tdrfstf`s fh Ds`adrf`afi @njf gi`tdrfi, usfhc pCJN pjismfk is i vd`tnr, cdhdtf`ijjy mnkflfdk gy rdstrf`tfnh dhzymds2 ihk tad cdhd waf`a fs rdspnhsfgjd lnr ljunrds`dh`d CLU (Crddh Ljunrds`dht Urntdfh). Ft ijsn `jirflfds Crim [d`ahfqud `nh`dpt, pnsftfvd ihk hdcitfvd Crim ihk tad ikvihticds nl bhnwfhc tad `nmpnsftfnh nl tad gi`tdrfij mdmgrihd. Uijivris @aivd? [æ`hf`i kn KHI Wd`nmgfhihtd, @iri`tdrístf`is Ldhntípf`is, Gi`tærfi,

Ds`adrf`afi @njf, Dhzfmis kd Wdstrfãàn, pCJN, Cdhd CLU, [æ`hf`i kd Crim, Crim Hdcitfvn, Crim Unsftfvn, Mdmgrihi Gi`tdrfihi.

Ds`nji Pd`uhkérfi Kr. Mihudj Cnmds kd Ijmdfki

Uécfhi 9 kd 93

Fhtrnkuãàn I `iki kfi qud pissi n muhkn oé hàn æ n mdsmn, i vfki trihslnrmi-sd, i Gfnjncfi rdfhvdhti-sd. Æ n `isn ki dhcdhairfi cdhætf`i, um dxdmpjn pdrldftn kd `nmn i ouhãàn kn pissir kn tdmpn `nm is pntdh`fijfkikds ki mdhtd aumihi `njignrim dm pdrldftn uhíssnhn. [nkns ns kfis hnvis kds`ngdrtis sàn ldftis, pnr dxpdrfmdhtiãàn, tdnrfi d prétf`i ngtïm-sd hnvis pntdh`fijfkikds `ipizds kd ijtdrir fkdfis prævfis d hdstd `isn tdstir mætnkns ijtdrhitfvns `nm n ngod`tfvn kd qudgrir girrdfris. Dm dspd`fij, n FUI[FMTU (Fhstftutn kd Uitnjncfi d Fmuhnjncfi Mnjd`ujir ki Thfvdrsfkikd kn Unrtn) mdrd`d sdr mdh`fnhikn pnfs sdrvd kd rdldrïh`fi hn `impn ki dhcdhairfi cdhætf`i pnr kds`ngdrtis sfchflf`itfvis d pfnhdfris, d timgæm pnr tdsds d irtfcns pugjf`ikns hdsti érdi. Tm kns pirçmdtrns dstukikns d rdjdvihtds kdhtrn ki dhcdhairfi cdhætf`i d pnstn dm prétf`i hi fki kns iutnrds kdstd pipdr in Jignritðrfn Igdrtn kdstd fhstftutn, sàn is i`tfvfkikds `nm gi`tærfis ljunrds`dhtds d i rdspd`tfvi mihfpujiãàn kn sdu KHI tdhkn dm vfsti rdspnhkdr ås sdcufhtds pdrcuhtis? @nmn fhtrnkuzfr um pjismíkdn humi gi`tærfi> Pdré pnssívdj mihfpujir n sdu KHI kd mnkn i tnrhé-ji mifs rdsfstdhtd i um ihtfgfðtf`n dspd`ílf`n> N ngod`tfvn ki i`tfvfkikd rdijfziki hn FUI[FMTU æ fcuijmdhtd pirtfjaikn pdjn ngod`tfvn kdstd kn`umdhtn `fdhtílf`n, ijircir `nhad`fmdhtns rdjitfvns i KHI rd`nmgfhihtd, gi`tærfis, ihtfgfðtf`ns d fhkuãàn kd hnvis `iri`tdrístf`is ldhntípf`is. Dsti i`tfvfkikd rdijfziki kdvfkn å npnrtuhfkikd prnpnr`fnhiki pdjn Jignritðrfn Igdrtn, pdrmftd umi rdjiãàn kfrd`ti `nm i kfs`fpjfhi kd Gfnjncfi pnr igrihcdr `nh`dftns tifs `nmn? prn`dssns kd mihfpujiãàn kd cdhds hum kikn nrcihfsmn (cdrijmdhtd lnri kn prn`dssn rdprnkutfvn hnrmij kdstd), fsnjimdhtn, mihfpujiãàn d fhtrnkuãàn kd KHI hum `aimikn `nrpn kd prnvi (nu anspdkdfrn), tdhkn `nmn ngod`tfvn i dxprdssàn kd um cdhd dspd`ílf`n prdvfimdhtd fkdhtflf`ikn d `itdcnrfzikn. I fmpnrtçh`fi tdðrf`i d prétf`i ki dxpdrfïh`fi rdijfziki, rdsfkfu hi npnrtuhfkikd kiki ins pirtf`fpihtds kd dxpjnrir d pûr dm prétf`i `nhtdÿkns ignrkikns hi kfs`fpjfhi issfm `nmn issfstfr in rdsujtikn kdssi mdsmi dxpdrfmdhtiãàn. N prdsdhtd pipdr dsté kfvfkfkn dm kuis pirtds prfh`fpifs? Uirtd F. Gi`tærfis Ljunrds`dhtds d Uirtd FF. [æ`hf`i kd Crim.

Ds`nji Pd`uhkérfi Kr. Mihudj Cnmds kd Ijmdfki

Uécfhi 7 kd 93

F. Gi`tærfis Ljunrds`dhtds Luhkimdhtns [dðrf`ns

Gi`tærfi Is gi`tærfis sàn prn`irfnhtds uhf`djujirds tdhkn, pnr fssn, umi `nhstftufãàn muftn sfmpjds. Pàn sdrds qud hàn iprdsdhtim nrcihdjns mdmgrihirds, hdm hÿ`jdn fhkfvfkuijfzikn, tdhkn n KHI kfspdrsn hn `ftnpjismi d hàn issn`fikn i afstnhis. Uiri ijæm ki mnjæ`uji kd KHI prfh`fpij, is gi`tærfis pnssudm ifhki pnrãôds kd KHI `fr`ujir dm `ikdfi kupji kdhnmfhikis Ujismíkdns. Dstds rdpjf`im-sd fhkdpdhkdhtdmdhtd ki mnjæ`uji kd KHI prfh`fpij d hàn sàn dssdh`fifs å sngrdvfvïh`fi ki gi`tærfi. Nrcihdjn

Luhãàn

Mdmgrihi @ftnpjismétf`i

@iptiãàn kd djdmdhtns hd`dssérfns d dx`rdãàn kd tnxfhis

Uirdkd @djujir

Wfcfkdz d Urntd`ãàn

@épsuji

Afkritiãàn Urntd`ãàn `nhtri Licn`ftnsd Mifnr ikdsàn i nutris `æjujis

Ufjf

Urdhkdr n Ijfmdhtn

Ljicdjn

Jn`nmnãàn

Wfgnssnmis

Píhtdsd Urntdf`i

Ujismíkdns

Md`ihfsmn kd Kdldsi

Hu`jdðfkd

Mitdrfij Cdhætf`n @nkflf`i is `iri`tdrístf`is ki `æjuji

Lfcuri 9 - Gi`tærfi

[æ`hf`i kn KHI rd`nmgfhihtd I [æ`hf`i kn KHI rd`nmgfhihtd æ utfjfziki dm dhcdhairfi cdhætf`i piri pnssfgfjftir qud kdtdrmfhiki dspæ`fd ikqufri `iri`tdrístf`is ldhntípf`is hàn fhitis in sdu cdhnmi. Æ fhtrnkuzfki umi pnrãàn kd KHI dxðcdhn prdvfimdhtd sdjd``fnhikn itrivæs kd dhzfmis kd rdstrfãàn. Dsti tæ`hf`i tdm um djdvikn pnkdr d`nhðmf`n pnfs pnssfgfjfti i prnkuãàn dm missi kd sugstçh`fis hd`dssérfis å sngrdvfvïh`fi (pntdh`fi tritimdhtns) kis quifs æ dxdmpjn i fhsujfhi, tàn hd`dssérfi piri fhsujfhnkdpdhkdhtds.

Ds`nji Pd`uhkérfi Kr. Mihudj Cnmds kd Ijmdfki

Uécfhi 0 kd 93

Uiri mnkflf`ir cdhdtf`imdhtd um nrcihfsmn æ hd`dssérfn ijtdrir tnkis is suis `æjujis,  oé qud tnkis djis pnssudm n cdhnmi kn fhkfvíkun. Pdhkn issfm, hdsti tæ`hf`i sàn utfjfzikns nrcihfsmns `nmn is gi`tærfis nu jdvdkuris kdvfkn å sui uhf`djujirfkikd d å mifnr li`fjfkikd kd mihfpujiãàn kn sdu `nhtdÿkn cdhætf`n pdji sui sfmpjf`fkikd, pdjn rdkuzfkn hÿmdrn kd hu`jdðtfkns qud n `nhstftudm d pdji sui répfki d dlf`iz rdprnkuãàn, nrfcfhihkn `jnhds.

Lfcuri 0 -Dsqudmitfziãàn ki i`ãàn kd umi Dhzfmi kd

Lfcuri 9 -Thfàn dhtrd n crupn lnslitn jfcikn in

`irgnhn 4  d n crupn afkrnxfji jfcikn in `irgnhn 0 kis mnjæ`ujis kd kdsnxfrfgnsd prdsdhtds hn KHI. ‷ 

‖

Wdstrf`ãàn (D`nWF)

Æ utfjfzikn um vd`tnr (mnjæ`uji `ipiz kd trihspnrtir n cdhd ds`njafkn piri i `æjuji anspdkdfri) qud `nhtæm n cdhd rd`njafkn ki mnjæ`uji kd KHI kiknri, hdstd `isn, n pjismíkdn. Imgns, pjismíkdn d mnjæ`uji kiknri, sàn tritikns `nm i mdsmi dhzfmi kd rdstrfãàn. Ns lricmdhtns kd rdstrfãàn (prnvdhfdhtds kn KHI kiknr), d ns pjismíkdns `nrtikns tïm dxtrdmfkikds dm `ikdfi sfmpjds (dxtrdmfkikds `ndsfvis). @njn`im-sd, pnstdrfnrmdhtd, ns lricmdhtns kd rdstrfãàn ouhtn ins pjismíkdns d is dxtrdmfkikds `ndsfvis kd imgns dmpirdjaim pnr `nmpjdmdhtirfkikd kd gisds, lnrmihkn pnhtds kd afkrncæhfn. Æ fhtrnkuzfki umi dhzfmi dspd`ílf`i, KHI jfcisd, qud dstigdjd`d i jfciãàn dhtrd is kuis `ikdfis sfmpjds (jfciãôds `nvijdhtd  ‐  lnslnkfæstdr), nrfcfhihkn umi hnvi mnjæ`uji kd KHI  ‐  rKHI (KHI rd`nmgfhihtd). N pjismíkdn rdsujtihtd kdstd prn`dssn kd ijtdriãàn ki sdquïh`fi hu`jdntíkf`i, icnri kdhnmfhikn pjismíkdn rd`nmgfhihtd, æ pnstn dm `nhti`tn `nm is `æjujis anspdkdfris (gi`tærfis). Dstd vd`tnr rd`nmgfhihtd æ ifhki pnrtiknr kd um cdhd qud jad `nhldrd rdsfstïh`fi i kdtdrmfhikn ihtfgfðtf`n piri pnstdrfnr sdjd`ãàn kis gi`tærfis trihslnrmikis. Is gi`tærfis qud fh`nrpnririm n pjismíkdn rd`nmgfhihtd fhf`fim i síhtdsd ki hnvi prntdíhi `nkflf`iki pdjn cdhd dxðcdhn fhtrnkuzfkn hn pjismíkdn. N rdsujtikn prdtdhkfkn sdré, dhtàn, i prnkuãàn dm missi kd kdtdrmfhiki gfnmnjæ`uji `nkflf`iki pdjn cdhd rd`njafkn ki mnjæ`uji kiknri.

Ds`nji Pd`uhkérfi Kr. Mihudj Cnmds kd Ijmdfki

Uécfhi ; kd 93

_nhi

@nkflf`i

Nrf (Nrfcfh)

Nrfcdm ki rdpjf`iãàn pjismíkf`i

Gji ( -ji`timisd cdhd)

Wdsfstïh`fi å impf`fjfhi (ihtfgfðtf`n)

CLU (Crddh Ljunrds`dht Urntdfh)

Píhtdsd ki prntdíhi qud `nhldrd ljunrds`ïh`fi

iri-@ (gfluh`tfnhij rdcujitnry prntdfh)

@nhtrnjn ki dxprdssàn kn CLU dm luhãàn ki prdsdhãi nu iusïh`fi kd Irigfhnsd hn mdfn

Ujismíkdn pCJN Æ itrivæs ki tæ`hf`i rdsumfkimdhtd kds`rfti i`fmi qud æ sfhtdtfzikn, jignritnrfijmdhtd, n pjismíkdn pCJN. I `nhstftufãàn kdstd pjismíkdn æ rdspnhsévdj pdji sui utfjfziãàn hi dxpdrfïh`fi ‚Gi`tærfis Ljunrds`dhtds” d kdsti`im-sd quitrn znhis prfh`fpifs. N pjismíkdn dm `iusi, pnstn dm `nhti`tn `nm is `æjujis anspdkdfris, fhf`firé i síhtdsd ki prntdíhi qud jads `nhldrd ljunrds`ïh`fi quihkn ngsdrvikis å juz ujtrivfnjdti. Ns prn`irfnhtds qud fhtdcriràn n pjismíkdn pCJN, hdstd `isn Ds`adrf`afi @njf , tnrhir-sd-àn rdsfstdhtds in ihtfgfðtf`n Impf`fjfhi (kdvfkn in cdhd gji), `iri`tdrístf`i qud hàn pnssufim hiturijmdhtd. Pdhkn issfm, dstd cdhd sdrvfré piri sdjd``fnhir hi `ujturi gi`tdrfihi, ns fhkfvíkuns qud fh`nrpnririm n pjismíkdn, dxpnhkn-ns å impf`fjfhi. Ipdhis ns qud sngrdvfvdm `nhtïm n vd`tnr mihfpujikn. Dsti sdjdãàn pdrmftd `drtflf`ir qud ipdhis is gi`tærfis mnkflf`ikis sd rdprnkuzdm, `jnhihkn n cdhd kd fhtdrdssd su`dssfvimdhtd.

pCJN

Lfcuri ; - Wdprdsdhtiãàn kn Ujismíkdn rd`nmgfhihtd pCJN

I znhi iri-@ æ rdspnhsévdj pdji rdcujiãàn ki dxprdssàn kn cdhd CLU, dstd ipdhis æ trihs`rftn hi prdsdhãi kd Irigfhnsd (kfssi`iríkdn). Æ um npdràn fhkutívdj, `nhstftuíkn pnr um npdriknr, um prnmntnr d trïs cdhds dstruturifs qud `nkflf`im i síhtdsd kis prntdíhis hd`dssérfis å kdcrikiãàn ki Irigfhnsd? n iriI, qud `nkflf`i i fsnmdrisd, n iriG qud `nkflf`i i rfgujn`fhisd d n iriK, qud `nkflf`i i rfgujnsd 4-lnslitn dpfmdrisd.

Ds`nji Pd`uhkérfi Kr. Mihudj Cnmds kd Ijmdfki

Uécfhi 4 kd 93

Npdràn Irigfhnsd Tm npdràn æ i uhfkikd rdcujitðrfi `nmpjdti kd um `nhouhtn kd cdhds icrupikns. Æ `nhstftuíkn pnr um cdhd rdcujiknr, rdspnhsévdj pdji síhtdsd kd umi prntdíhi rdprdssnri2 um cdhd npdriknr, rdspnhsévdj pdji lfxiãàn ki prntdíhi rdprdssnri in npdràn2 um cdhd prnmntnr, rdspnhsévdj pdji lfxiãàn ki WHI pnjfmdrisd2 d um `nhouhtn kd cdhds dstruturifs, rdspnhsévdfs pdji `nkflf`iãàn kns imfhné`fkns qud, ouhtns, lnrmiràn is prntdíhis rdspnhsévdfs pdji kdcrikiãàn ki irigfhnsd. Pð æ hd`dssérfi i síhtdsd kis prntdíhis kdcrikiknris kn kfssi`iríkdn sd dstd dstfvdr prdsdhtd hn mdfn. @isn `nhtrérfn sdré um kdspdrkí`fn kd dhdrcfi. Pdhkn qud is gi`tærfis ipdhis prnkuzdm I[U itrivæs ki Ldrmdhtiãàn (n gijihãn lfhij sàn kuis mnjæ`ujis kd I[U) hàn pnkdm lizdr kdspdrkí`fns. Hi iusïh`fi kd irigfhnsd hn mdfn i prntdíhi rdprdssnri æ sfhtdtfziki hi lnrmi i`tfvi, lfxi-sd in npdriknr fh`ipi`ftihkn i lfxiãàn ki WHI pnjfmdrisd. Hàn aé trihs`rfãàn kns cdhds dstruturifs, `nhsdqudhtdmdhtd hàn aé trikuãàn hdm síhtdsd prntdf`i.

Lfcuri 4 - Dsqudmi kn Npdràn Irigfhnsd F

Ds`nji Pd`uhkérfi Kr. Mihudj Cnmds kd Ijmdfki

Uécfhi @nmn tnrhé-ji rdsfstdhtd i kdtdrmfhikn ihtfgfðtf`n> I i`tfvfkikd prnpnsti pdrmftd, rd`nrrdhkn å tæ`hf`i kn KHI rd`nmgfhihtd, i mihfpujiãàn kd gi`tærfis d i sui `nmpiriãàn `nm gi`tærfis hàn mihfpujikis. Ipðs tnki i fhlnrmiãàn tdðrf`i rd`njafki d pdr`dgfki, `rfirim-sd is sdcufhtds afpðtdsds? Is gi`tærfis cdhdtf`imdhtd mnkflf`ikis, nu sdoi, qud fh`nrpnririm n pjismíkdn rd`nmgfhihtd, dmftdm ljunrds`ïh`fi quihkn ngsdrvikis å juz Tjtri \fnjdti sd dstfvdrdm hi prdsdhãi kd Irigfhnsd2 dxpnstis in ihtfgfðtf`n (Impf`fjfhi), dstis gi`tærfis sngrdvfvdm. Is gi`tærfis qud hàn lnrim pnstis dm `nhti`tn `nm n pjismíkdn rd`nmgfhihtd, hàn n fh`nrpnririm, jncn hàn sngrdvfvdràn dm mdfns kd `ujturi nhkd dstdoi prdsdhtd n ihtfgfðtf`n Impf`fjfhi. Ki mdsmi lnrmi, hàn iprdsdhtiràn ljunrds`ïh`fi kdvfkn å iusïh`fi kn CLU.

@nm n ngod`tfvn kd vijfkir dstis afpðtdsds lnf dxd`utiki i dxpdrfïh`fi ki sdcufhtd mihdfri. 9. Mf`rnpfpdtiãàn kd 745 j kd Pnjuãàn kd [rihslnrmiãàn (@i@j7) piri n mf`rntugn. 7. Wd`njai kd umi pdqudhi pnrãàn kd Gi`tærfis ki dspæ`fd Ds`adrf`afi @njf   `nm i Ihsi kds`irtévdj d fhsdrãàn kis mdsmis hn mf`rntugn. 0. Wd`njai kd umi pdjí`uji kd pjismíkdns pCJN `nm nutri Ihsi kds`irtévdj d fhsdrãàn kns mdsmns hn mf`rntugn. ;. Fh`ugiãàn kn mf`rntugn dm cdjn, ipðs kdvfki fkdhtflf`iãàn 9, kurihtd iprnxfmikimdhtd 0 mfhutns. 9

Pdhkn qud n mf`rntugn `nhtfhai pjismíkdns pCJN lnf fkdhtflf`ikn `nm um + (=mifs), tnkns ns qud hàn `nhtfhaim KHI pjismíkf`n lnrim fkdhtflf`ikns `nm um  ‐ (=mdhns).

Ds`nji Pd`uhkérfi Kr. Mihudj Cnmds kd Ijmdfki

Uécfhi 6 kd 93

4. [rihsldrïh`fi kn mf`rntugn piri n Gihan Mirfi i ;7 n@ kurihtd iprnxfmikimdhtd 9 mfhutn.