{"id":37174,"date":"2019-10-31T22:16:11","date_gmt":"2019-10-31T19:16:11","guid":{"rendered":"https:\/\/prohoster.info\/blog\/umer-nobelevskij-laureat-keri-mullis-izobretatel-polimeraznoj-tsepnoj-reaktsii-dnk\/"},"modified":"2019-10-31T22:16:11","modified_gmt":"2019-10-31T19:16:11","slug":"umer-nobelevskij-laureat-keri-mullis-izobretatel-polimeraznoj-tsepnoj-reaktsii-dnk","status":"publish","type":"post","link":"https:\/\/prohoster.info\/cs\/blog\/news\/umer-nobelevskij-laureat-keri-mullis-izobretatel-polimeraznoj-tsepnoj-reaktsii-dnk","title":{"rendered":"Zem\u0159el nositel Nobelovy ceny Kary Mullis \u2013 vyn\u00e1lezce polymer\u00e1zov\u00e9 \u0159et\u011bzov\u00e9 reakce DNA","gt_translate_keys":[{"key":"rendered","format":"text"}]},"content":{"rendered":"

\"Zem\u0159el V Kalifornii ve v\u011bku 74 let zem\u0159el americk\u00fd nositel Nobelovy ceny za chemii Kary Mullis. Podle slov jeho man\u017eelky, smrt nastala 7. srpna. P\u0159\u00ed\u010dina \u2014 srde\u010dn\u00ed a dechov\u00e1 nedostate\u010dnost zp\u016fsoben\u00e1 pneumoni\u00ed.<\/p>\n

O tom, jak\u00fd p\u0159\u00ednos m\u011bl pro bioch\u00e9mii a za co z\u00edskal Nobelovu cenu, n\u00e1m pov\u00ed s\u00e1m James Watson \u2014 objevitel molekuly DNA. <\/p>\n

\u00daryvek z knihy Jamese Watsona, Andrewa Berryho, Kevina Davise <\/p>\n

DNA. Historie genetick\u00e9 revoluce<\/h2>\n

Kapitola 7. Genom \u010dlov\u011bka. Sc\u00e9n\u00e1\u0159 \u017eivota<\/h3>\n

\n<\/a><\/noindex>
\n\u2026
\nPolymer\u00e1zov\u00e1 \u0159et\u011bzov\u00e1 reakce (PCR) byla vynalezena v roce 1983 bioch\u00e9mi\u010dkou Kary Mullisem, kter\u00fd pracoval ve spole\u010dnosti Cetus. Objev t\u00e9to reakce byl velmi v\u00fdjime\u010dn\u00fd. Pozd\u011bji Mullis vzpom\u00ednal: \u201eJednoho p\u00e1te\u010dn\u00edho ve\u010dera v dubnu 1983 m\u011b jakoby osv\u00edtilo. \u0158\u00eddil jsem, proj\u00ed\u017ed\u011bl po osv\u011btlen\u00e9 k\u0159ivolak\u00e9 horsk\u00e9 silnici v severn\u00ed Kalifornii, kraji sekvojov\u00fdch les\u016f.\u201c Je p\u016fsobiv\u00e9, \u017ee pr\u00e1v\u011b v takov\u00e9 situaci ho nav\u0161t\u00edvila inspirace. A nen\u00ed to jen t\u00edm, \u017ee na severu Kalifornie jsou zvl\u00e1\u0161tn\u00ed silnice, kter\u00e9 podporuj\u00ed osv\u00edcen\u00ed; jeho p\u0159\u00edtel jednou vid\u011bl, jak se Mullis bezstarostn\u011b \u0159\u00edtil po namrzl\u00e9 silnici s obousm\u011brn\u00fdm provozem a to ho v\u016fbec nevzru\u0161ovalo. P\u0159\u00edtel uvedl pro New York Times: \u201eMullisovi se zd\u00e1lo, \u017ee zahyne, kdy\u017e naraz\u00ed do sekvoje. Proto se ni\u010deho neboj\u00ed za volantem, pokud pod\u00e9l silnice nerostou sekvojovce.\u201c P\u0159\u00edtomnost sekvoj\u016f pod\u00e9l silnice nutila Mullise soust\u0159edit se a\u2026 a zde je to, osv\u00edcen\u00ed. Za sv\u016fj objev v roce 1993 z\u00edskal Mullis Nobelovu cenu za chemii a od t\u00e9 doby se jeho chov\u00e1n\u00ed stalo je\u0161t\u011b podivn\u011bj\u0161\u00edm. Nap\u0159\u00edklad je zast\u00e1ncem revizionistick\u00e9 teorie, \u017ee AIDS nen\u00ed spojen s HIV, co\u017e zna\u010dn\u011b po\u0161kodilo jeho vlastn\u00ed reputaci a br\u00e1nilo l\u00e9ka\u0159\u016fm.<\/p>\n

PCR je pom\u011brn\u011b jednoduch\u00e1 reakce. K jej\u00edmu proveden\u00ed pot\u0159ebujeme dva chemicky syntetizovan\u00e9 primery, komplement\u00e1rn\u00ed k opa\u010dn\u00fdm konc\u016fm r\u016fzn\u00fdch \u0159et\u011bzc\u016f po\u017eadovan\u00e9ho fragmentu DNA. Primery jsou kr\u00e1tk\u00e9 \u00faseky jedno\u0159et\u011bzcov\u00e9 DNA, ka\u017ed\u00fd p\u0159ibli\u017en\u011b 20 p\u00e1r\u016f b\u00e1z\u00ed dlouh\u00fd. Specifi\u010dnost primer\u016f spo\u010d\u00edv\u00e1 v tom, \u017ee odpov\u00eddaj\u00ed \u00fasek\u016fm DNA, kter\u00e9 je t\u0159eba amplifikovat, tedy DNA matrici. <\/p>\n

\"Zem\u0159el<\/a><\/noindex>
\n(Obr\u00e1zek je klikac\u00ed) Kary Mullis, vyn\u00e1lezce PCR<\/i><\/p>\n

Specifi\u010dnost PCR spo\u010d\u00edv\u00e1 v tvorb\u011b komplement\u00e1rn\u00edch komplex\u016f mezi \u0161ablonou a primery, kr\u00e1tk\u00fdmi syntetick\u00fdmi oligonucl\u00e9otidy. Ka\u017ed\u00fd z primer\u016f je komplement\u00e1rn\u00ed k jednomu z \u0159et\u011bzc\u016f dvouvl\u00e1knov\u00e9 \u0161ablony a vymezuje za\u010d\u00e1tek a konec amplifikovan\u00e9 oblasti. Skute\u010dn\u00e1 \u201e\u0161ablona\u201c p\u0159edstavuje celkov\u00fd genom, a na\u0161\u00edm c\u00edlem je extrahovat z n\u00ed fragmenty, kter\u00e9 n\u00e1s zaj\u00edmaj\u00ed. K tomu se dvouvl\u00e1knov\u00e1 DNA \u0161ablona zah\u0159\u00edv\u00e1 na 95 \u00b0C na n\u011bkolik minut, aby se \u0159et\u011bzce DNA odd\u011blily. Tato f\u00e1ze se naz\u00fdv\u00e1 denaturace, proto\u017ee doch\u00e1z\u00ed k rozru\u0161en\u00ed vod\u00edkov\u00fdch vazeb mezi dv\u011bma \u0159et\u011bzci DNA. Jakmile se \u0159et\u011bzce odd\u011bl\u00ed, teplota se sn\u00ed\u017e\u00ed, aby se primery mohly nav\u00e1zat na jedno\u0159et\u011bzcovou \u0161ablonu. DNA polymer\u00e1za za\u010d\u00edn\u00e1 replikaci DNA t\u00edm, \u017ee se v\u00e1\u017ee na \u00fasek \u0159et\u011bzce nukleotid\u016f. Enzym DNA polymer\u00e1za replikovuje \u0161ablonov\u00fd \u0159et\u011bzec, p\u0159i\u010dem\u017e pou\u017e\u00edv\u00e1 primer jako start\u00e9r pro kop\u00edrov\u00e1n\u00ed. V d\u016fsledku prvn\u00edho cyklu z\u00edsk\u00e1v\u00e1me opakovan\u00e9 sekven\u010dn\u00ed dvojn\u00e1soben\u00ed ur\u010dit\u00e9ho \u00faseku DNA. Pot\u00e9 tuto proceduru opakujeme. Po ka\u017ed\u00e9m cyklu z\u00edsk\u00e1v\u00e1me c\u00edlovou oblast ve dvojn\u00e1sobn\u00e9m mno\u017estv\u00ed. Po dvaceti p\u011bti cyklech PCR (tj. za m\u00e9n\u011b ne\u017e dv\u011b hodiny) m\u00e1me po\u017eadovan\u00fd \u00fasek DNA v mno\u017estv\u00ed, kter\u00e9 je 225kr\u00e1t vy\u0161\u0161\u00ed ne\u017e v\u00fdchoz\u00ed (tj. amplifikovali jsme jej p\u0159ibli\u017en\u011b 34 milionkr\u00e1t). Ve skute\u010dnosti jsme na vstupu m\u011bli sm\u011bs primer\u016f, \u0161ablonov\u00e9 DNA, enzymu DNA polymer\u00e1zy a voln\u00fdch b\u00e1z\u00ed A, C, G a T, p\u0159i\u010dem\u017e mno\u017estv\u00ed specifick\u00e9ho produktu reakce (omezen\u00e9ho primery) roste exponenci\u00e1ln\u011b, zat\u00edmco mno\u017estv\u00ed \u201edlouh\u00fdch\u201c kopi\u00ed DNA line\u00e1rn\u011b, tak\u017ee v produktech reakce dominuje.<\/p>\n

\"Zem\u0159el
\nAmplifikace po\u017eadovan\u00e9ho \u00faseku DNA: polymer\u00e1zov\u00e1 \u0159et\u011bzov\u00e1 reakce<\/i><\/p>\n

Na za\u010d\u00e1tku pou\u017e\u00edv\u00e1n\u00ed PCR byla hlavn\u00edm probl\u00e9mem n\u00e1sleduj\u00edc\u00ed situace: po ka\u017ed\u00e9m cyklu oh\u0159evu a chlazen\u00ed bylo nutn\u00e9 p\u0159id\u00e1vat do reak\u010dn\u00ed sm\u011bsi DNA polymer\u00e1zu, proto\u017ee p\u0159i teplot\u011b 95 \u00b0C inaktivovala. Bylo tedy nutn\u00e9 ji znovu p\u0159id\u00e1vat p\u0159ed ka\u017ed\u00fdm z 25 cykl\u016f. Veden\u00ed reakce bylo relativn\u011b neefektivn\u00ed, vy\u017eadovalo hodn\u011b \u010dasu a enzym byl pom\u011brn\u011b drah\u00fd. Na\u0161t\u011bst\u00ed p\u0159i\u0161la na pomoc sama p\u0159\u00edroda. Mnoho \u017eivo\u010dich\u016f se c\u00edt\u00ed pohodln\u011b p\u0159i teplot\u00e1ch v\u00fdrazn\u011b vy\u0161\u0161\u00edch ne\u017e 37 \u00b0C. A pro\u010d je pro n\u00e1s \u010d\u00edslo 37 \u00b0C tak d\u016fle\u017eit\u00e9? D\u011bje se tak proto, \u017ee tato teplota je optim\u00e1ln\u00ed pro E. coli, ze kter\u00e9 byl p\u016fvodn\u011b z\u00edsk\u00e1n enzym polymer\u00e1zy pro PCR. V p\u0159\u00edrod\u011b se vyskytuj\u00ed mikroorganismy, jejich\u017e proteiny se b\u011bhem milion\u016f let p\u0159irozen\u00e9ho v\u00fdb\u011bru staly odoln\u011bj\u0161\u00edmi v\u016f\u010di vysok\u00fdm teplot\u00e1m. Bylo navr\u017eeno vyu\u017e\u00edt DNA polymer\u00e1zy z termofiln\u00edch bakteri\u00ed. Tyto enzymy se uk\u00e1zaly jako termostabiln\u00ed a byly schopny vydr\u017eet mnoho cykl\u016f reakce. Jejich pou\u017eit\u00ed usnadnilo a automatizovalo prov\u00e1d\u011bn\u00ed PCR. Jedna z prvn\u00edch termostabiln\u00edch DNA polymer\u00e1z byla izolov\u00e1na z bakterie Thermus aquaticus \u017eij\u00edc\u00ed v hork\u00fdch pramenech Yellowstone, a byla nazv\u00e1na Taq polymer\u00e1zou.<\/p>\n

PCR se rychle stala hlavn\u00edm pracovn\u00edm n\u00e1strojem projektu \u201eLidsk\u00fd genom\u201c. Celkov\u011b proces nezm\u011bnil od n\u00e1vrhu Mullise, jen byl automatizov\u00e1n. ji\u017e jsme nebyli z\u00e1visl\u00ed na skupin\u011b ne\u0161ikovn\u00fdch postgradu\u00e1ln\u00edch student\u016f, kte\u0159\u00ed pe\u010dliv\u011b p\u0159elivali kapky tekutiny do plastov\u00fdch zkumavek. V modern\u00edch laborato\u0159\u00edch, kter\u00e9 prov\u00e1d\u011bj\u00ed molekul\u00e1rn\u011b genetick\u00e1 vy\u0161et\u0159en\u00ed, tuto pr\u00e1ci vykon\u00e1vaj\u00ed robotizovan\u00e9 linky. PCR roboti zapojen\u00ed do tak rozs\u00e1hl\u00e9ho sekvenovac\u00edho projektu, jako je \u201eLidsk\u00fd genom\u201c, ne\u00fanavn\u011b pracuj\u00ed s obrovsk\u00fdmi objemy teplotn\u011b odoln\u00e9 polymer\u00e1zy. N\u011bkte\u0159\u00ed v\u011bdci pracuj\u00edc\u00ed na projektu \u201eLidsk\u00fd genom\u201c byli rozho\u0159\u010deni nep\u0159im\u011b\u0159en\u011b vysok\u00fdmi poplatky, kter\u00e9 p\u0159id\u00e1v\u00e1 k n\u00e1klad\u016fm na spot\u0159ebn\u00ed materi\u00e1l majitel patentu na PCR, evropsk\u00fd farmaceutick\u00fd gigant Hoffmann-LaRoche.<\/p>\n

Dal\u0161\u00edm \"pohybuj\u00edc\u00edm se prvkem\" byla sama metoda sekvenov\u00e1n\u00ed DNA. Chemick\u00fd z\u00e1klad t\u00e9to metody ji\u017e nebyl novinkou: Mezist\u00e1tn\u00ed projekt \"Lidsk\u00fd genom\" (HGP) si osvojil stejnou vysoce sofistikovanou metodu, kterou vyvinul Fred Sanger ji\u017e v polovin\u011b 70. let. Inovace v\u0161ak spo\u010d\u00edvala v rozsahu a m\u00ed\u0159e automatizace, kter\u00fdch se poda\u0159ilo dos\u00e1hnout p\u0159i sekvenov\u00e1n\u00ed.<\/p>\n

Automatick\u00e9 sekvenov\u00e1n\u00ed bylo p\u016fvodn\u011b vyv\u00edjeno v laborato\u0159i Leeho Huda na Kalifornsk\u00e9m technologick\u00e9m institutu. Lee dokon\u010dil \u0161kolu ve st\u00e1t\u011b Montana a hr\u00e1l americk\u00fd fotbal na pozici \u00fato\u010dn\u00edka; d\u00edky Hudo t\u00fdm n\u011bkolikr\u00e1t vyhr\u00e1l st\u00e1tn\u00ed \u0161ampion\u00e1t. Dovednosti t\u00fdmov\u00e9 spolupr\u00e1ce se mu hodily i v jeho v\u011bdeck\u00e9 kari\u00e9\u0159e. V Huddov\u011b laborato\u0159i pracovala pestr\u00e1 skupina chemik\u016f, biolog\u016f a in\u017een\u00fdr\u016f, a brzy se jeho laborato\u0159 stala l\u00eddrem v technologick\u00fdch inovac\u00edch.<\/p>\n

Ve skute\u010dnosti metod automatick\u00e9ho sekvenov\u00e1n\u00ed vynalezli Lloyd Smith a Mike Hunkapiller. Mike Hunkapiller, kter\u00fd tehdy pracoval v laboratoriu Huda, oslovil Lloyda Smitha s n\u00e1vrhem na vylep\u0161enou metodu sekvenov\u00e1n\u00ed, ve kter\u00e9 by byly b\u00e1ze ka\u017ed\u00e9ho typu barveny ka\u017ed\u00e1 jinou barvou. Tato my\u0161lenka mohla \u010dty\u0159ikr\u00e1t zv\u00fd\u0161it efektivitu Sangerova procesu. P\u0159i sekvenov\u00e1n\u00ed podle Sangerova postupu se v ka\u017ed\u00e9 ze \u010dty\u0159 zkumavek (podle po\u010dtu b\u00e1z\u00ed) za \u00fa\u010dasti DNA polymer\u00e1zy vytv\u00e1\u0159\u00ed jedine\u010dn\u00e1 sada oligonukleotid\u016f r\u016fzn\u00fdch d\u00e9lek, zahrnuj\u00edc\u00edch primingovou sekvenci. Pot\u00e9 byly do zkumavek p\u0159id\u00e1ny formamid pro rozpojen\u00ed \u0159et\u011bzc\u016f a provedl se elektrofor\u00e9za v polyakrylamidov\u00e9m gelu na \u010dty\u0159ech dr\u00e1h\u00e1ch. Ve variant\u011b Smitha a Hunkapillera se dideoxynukleotidy ozna\u010duj\u00ed \u010dty\u0159mi r\u016fzn\u00fdmi barvivy a PCR se prov\u00e1d\u00ed v jedn\u00e9 zkumavce. Pot\u00e9 b\u011bhem elektrofor\u00e9zy v polyakrylamidov\u00e9m gelu laserov\u00fd paprsek na ur\u010dit\u00e9m m\u00edst\u011b gelu aktivuje \u010dinnost barviv, a detektor ur\u010duje, kter\u00fd nukleotid v sou\u010dasnosti migruje p\u0159es gel. Zpo\u010d\u00e1tku byl Smith nalad\u011bn pesimisticky - ob\u00e1val se, \u017ee pou\u017eit\u00ed extr\u00e9mn\u011b mal\u00fdch d\u00e1vek barviva povede k tomu, \u017ee nukleotidov\u00e9 \u00faseky budou nerozli\u0161iteln\u00e9. Nicm\u00e9n\u011b, s excelentn\u00ed znalost\u00ed laserov\u00fdch technologi\u00ed brzy na\u0161el \u0159e\u0161en\u00ed, kdy vyu\u017eil speci\u00e1ln\u00ed fluorescen\u010dn\u00ed barviva, kter\u00e1 fluoreskuj\u00ed pod vlivem laserov\u00e9ho z\u00e1\u0159en\u00ed.<\/p>\n

\"Zem\u0159el
\n(Pln\u00e1 verze po kliknut\u00ed<\/a><\/noindex> \u2014 4,08 MB) Mal\u00fdm p\u00edsmem: sekvence DNA, kterou dek\u00f3doval automatick\u00fd sekven\u00e1tor, z\u00edskan\u00e1 z apar\u00e1tu pro automatick\u00e9 sekvenov\u00e1n\u00ed. Ka\u017ed\u00e9 barv\u011b odpov\u00edd\u00e1 jedna ze \u010dty\u0159 b\u00e1z\u00ed<\/i> <\/p>\n

V klasick\u00e9 variant\u011b metody Sangerovy jedna z \u0159et\u011bzc\u016f analyzovan\u00e9 DNA slou\u017e\u00ed jako matice pro synt\u00e9zu komplement\u00e1rn\u00edho \u0159et\u011bzce enzymem DNA polymer\u00e1zou, pot\u00e9 se sekvence fragment\u016f DNA t\u0159\u00edd\u00ed v gelu podle velikosti. Ka\u017ed\u00fd fragment, kter\u00fd je sou\u010d\u00e1st\u00ed DNA b\u011bhem synt\u00e9zy a kter\u00fd umo\u017e\u0148uje n\u00e1slednou vizualizaci produkt\u016f reakce, je ozna\u010den fluorescen\u010dn\u00edm barvivem odpov\u00eddaj\u00edc\u00edm termin\u00e1ln\u00edmu z\u00e1kladu (o tom bylo zm\u00edn\u011bno na str. 124); t\u00edm p\u00e1dem bude fluorescen\u010dn\u00ed sign\u00e1l tohoto fragmentu identifik\u00e1torem pro dan\u00fd z\u00e1klad. Pot\u00e9 zb\u00fdv\u00e1 pouze prov\u00e9st detekci a vizualizovat produkty reakce. V\u00fdsledky se analyzuj\u00ed pomoc\u00ed po\u010d\u00edta\u010de a prezentuj\u00ed se ve form\u011b sekvence r\u016fznobarevn\u00fdch hrot\u016f, kter\u00e9 odpov\u00eddaj\u00ed \u010dty\u0159em nukleotid\u016fm. D\u00e1le je informace p\u0159ed\u00e1na p\u0159\u00edmo do informa\u010dn\u00edho syst\u00e9mu po\u010d\u00edta\u010de, co\u017e eliminuje \u010dasov\u011b n\u00e1ro\u010dn\u00fd a n\u011bkdy mu\u010div\u00fd proces zad\u00e1v\u00e1n\u00ed dat, kter\u00fd zna\u010dn\u011b komplikoval sekvenov\u00e1n\u00ed.<\/p>\n

\u00bb Podrobnosti o knize naleznete na webov\u00fdch str\u00e1nk\u00e1ch vydavatelstv\u00ed<\/a><\/noindex>
\n\u00bb Obsah<\/a><\/noindex>
\n\u00bb \u00daryvek<\/a><\/noindex><\/p>\n

Pro u\u017eivatele Habr sleva 25 % s kup\u00f3nem \u2014 PCR<\/b>
\n
Zdroj: habr.com<\/a><\/p>","protected":false,"gt_translate_keys":[{"key":"rendered","format":"html"}]},"excerpt":{"rendered":"

\u0412 \u041a\u0430\u043b\u0438\u0444\u043e\u0440\u043d\u0438\u0438 \u0432 \u0432\u043e\u0437\u0440\u0430\u0441\u0442\u0435 74 \u043b\u0435\u0442 \u0443\u043c\u0435\u0440 \u0430\u043c\u0435\u0440\u0438\u043a\u0430\u043d\u0441\u043a\u0438\u0439 \u043d\u043e\u0431\u0435\u043b\u0435\u0432\u0441\u043a\u0438\u0439 \u043b\u0430\u0443\u0440\u0435\u0430\u0442 \u043f\u043e \u0445\u0438\u043c\u0438\u0438 \u041a\u044d\u0440\u0438 \u041c\u0443\u043b\u043b\u0438\u0441. \u041f\u043e \u0441\u043b\u043e\u0432\u0430\u043c \u0435\u0433\u043e \u0441\u0443\u043f\u0440\u0443\u0433\u0438, \u0441\u043c\u0435\u0440\u0442\u044c \u043d\u0430\u0441\u0442\u0443\u043f\u0438\u043b\u0430 7 \u0430\u0432\u0433\u0443\u0441\u0442\u0430. \u041f\u0440\u0438\u0447\u0438\u043d\u0430 \u2014 \u0441\u0435\u0440\u0434\u0435\u0447\u043d\u0430\u044f \u0438 \u0434\u044b\u0445\u0430\u0442\u0435\u043b\u044c\u043d\u0430\u044f \u043d\u0435\u0434\u043e\u0441\u0442\u0430\u0442\u043e\u0447\u043d\u043e\u0441\u0442\u044c \u0438\u0437-\u0437\u0430 \u043f\u043d\u0435\u0432\u043c\u043e\u043d\u0438\u0438. \u041e \u0442\u043e\u043c, \u043a\u0430\u043a\u043e\u0439 \u0432\u043a\u043b\u0430\u0434 \u043e\u043d \u0432\u043d\u0435\u0441 \u0432 \u0431\u0438\u043e\u0445\u0438\u043c\u0438\u044e \u0438 \u0437\u0430 \u0447\u0442\u043e \u043f\u043e\u043b\u0443\u0447\u0438\u043b \u041d\u043e\u0431\u0435\u043b\u0435\u0432\u0441\u043a\u0443\u044e \u043f\u0440\u0435\u043c\u0438\u044e, \u043d\u0430\u043c \u0440\u0430\u0441\u0441\u043a\u0430\u0436\u0435\u0442 \u0441\u0430\u043c \u0414\u0436\u0435\u0439\u043c\u0441 \u0423\u043e\u0442\u0441\u043e\u043d \u2014 \u043f\u0435\u0440\u0432\u043e\u043e\u0442\u043a\u0440\u044b\u0432\u0430\u0442\u0435\u043b\u044c \u043c\u043e\u043b\u0435\u043a\u0443\u043b\u044b \u0414\u041d\u041a. \u041e\u0442\u0440\u044b\u0432\u043e\u043a \u0438\u0437 […]<\/p>\n","protected":false,"gt_translate_keys":[{"key":"rendered","format":"html"}]},"author":1,"featured_media":27873,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[702],"tags":[],"class_list":["post-37174","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-news"],"aioseo_notices":[],"aioseo_head":"\n\t\t\n\t\n\t\n\t\n\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\n","aioseo_head_json":{"title":"\ud83e\udd47Zem\u0159el nositel Nobelovy ceny Kary Mullis \u2014 vyn\u00e1lezce polymer\u00e1zov\u00e9 \u0159et\u011bzov\u00e9 reakce DNA | ProHoster","description":"V Kalifornii.","canonical_url":"https:\/\/prohoster.info\/cs\/blog\/news\/umer-nobelevskij-laureat-keri-mullis-izobretatel-polimeraznoj-tsepnoj-reaktsii-dnk","robots":"max-image-preview:large","keywords":"","webmasterTools":{"miscellaneous":""},"schema":null,"og:locale":"cs_CZ","og:site_name":"ProHoster | \u041a\u0443\u043f\u0438\u0442\u044c \u043d\u0430\u0434\u0435\u0436\u043d\u044b\u0439 \u0445\u043e\u0441\u0442\u0438\u043d\u0433 \u0434\u043b\u044f \u0441\u0430\u0439\u0442\u043e\u0432 \u0441 \u0437\u0430\u0449\u0438\u0442\u043e\u0439 \u043e\u0442 DDoS, VPS VDS \u0441\u0435\u0440\u0432\u0435\u0440\u044b","og:type":"article","og:title":"\ud83e\udd47\u0423\u043c\u0435\u0440 \u043d\u043e\u0431\u0435\u043b\u0435\u0432\u0441\u043a\u0438\u0439 \u043b\u0430\u0443\u0440\u0435\u0430\u0442 \u041a\u044d\u0440\u0438 \u041c\u0443\u043b\u043b\u0438\u0441 \u2014 \u0438\u0437\u043e\u0431\u0440\u0435\u0442\u0430\u0442\u0435\u043b\u044c \u043f\u043e\u043b\u0438\u043c\u0435\u0440\u0430\u0437\u043d\u043e\u0439 \u0446\u0435\u043f\u043d\u043e\u0439 \u0440\u0435\u0430\u043a\u0446\u0438\u0438 \u0414\u041d\u041a | ProHoster","og:description":"\u0412 \u041a\u0430\u043b\u0438\u0444\u043e\u0440\u043d\u0438\u0438.","og:url":"https:\/\/prohoster.info\/cs\/blog\/news\/umer-nobelevskij-laureat-keri-mullis-izobretatel-polimeraznoj-tsepnoj-reaktsii-dnk","og:image":"https:\/\/prohoster.info\/wp-content\/uploads\/2021\/11\/logo-350.jpg","og:image:secure_url":"https:\/\/prohoster.info\/wp-content\/uploads\/2021\/11\/logo-350.jpg","og:image:width":350,"og:image:height":350,"article:published_time":"2019-10-31T19:16:11+00:00","article:modified_time":"2019-10-31T19:16:11+00:00","article:publisher":"https:\/\/www.facebook.com\/prohoster","article:author":"https:\/\/www.facebook.com\/prohoster"},"aioseo_meta_data":{"post_id":"37174","title":null,"description":null,"keywords":null,"keyphrases":null,"primary_term":null,"canonical_url":null,"og_title":null,"og_description":null,"og_object_type":"default","og_image_type":"default","og_image_url":null,"og_image_width":null,"og_image_height":null,"og_image_custom_url":null,"og_image_custom_fields":null,"og_video":null,"og_custom_url":null,"og_article_section":null,"og_article_tags":null,"twitter_use_og":false,"twitter_card":"default","twitter_image_type":"default","twitter_image_url":null,"twitter_image_custom_url":null,"twitter_image_custom_fields":null,"twitter_title":null,"twitter_description":null,"schema":{"blockGraphs":[],"customGraphs":[],"default":{"data":{"Article":[],"Course":[],"Dataset":[],"FAQPage":[],"Movie":[],"Person":[],"Product":[],"ProductReview":[],"Car":[],"Recipe":[],"Service":[],"SoftwareApplication":[],"WebPage":[]},"graphName":"","isEnabled":true},"graphs":[]},"schema_type":null,"schema_type_options":null,"pillar_content":false,"robots_default":true,"robots_noindex":false,"robots_noarchive":false,"robots_nosnippet":false,"robots_nofollow":false,"robots_noimageindex":false,"robots_noodp":false,"robots_notranslate":false,"robots_max_snippet":null,"robots_max_videopreview":null,"robots_max_imagepreview":"large","priority":null,"frequency":null,"local_seo":null,"seo_analyzer_scan_date":"2026-01-22 06:26:23","breadcrumb_settings":null,"limit_modified_date":false,"reviewed_by":null,"ai":null,"created":"2021-03-01 01:32:13","updated":"2026-01-22 06:26:23","focus_keyword":null,"additional_keywords":null,"truseo_locale":null},"gt_translate_keys":[{"key":"link","format":"url"}],"_links":{"self":[{"href":"https:\/\/prohoster.info\/cs\/wp-json\/wp\/v2\/posts\/37174","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/prohoster.info\/cs\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/prohoster.info\/cs\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/prohoster.info\/cs\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/prohoster.info\/cs\/wp-json\/wp\/v2\/comments?post=37174"}],"version-history":[{"count":0,"href":"https:\/\/prohoster.info\/cs\/wp-json\/wp\/v2\/posts\/37174\/revisions"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/prohoster.info\/cs\/wp-json\/wp\/v2\/media\/27873"}],"wp:attachment":[{"href":"https:\/\/prohoster.info\/cs\/wp-json\/wp\/v2\/media?parent=37174"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/prohoster.info\/cs\/wp-json\/wp\/v2\/categories?post=37174"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/prohoster.info\/cs\/wp-json\/wp\/v2\/tags?post=37174"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}