{"id":107119,"date":"2026-05-22T00:16:18","date_gmt":"2026-05-21T22:16:18","guid":{"rendered":"https:\/\/www.infokuryr.cz\/n\/?p=107119"},"modified":"2026-05-21T14:18:01","modified_gmt":"2026-05-21T12:18:01","slug":"v-roce-2008-laborator-baric-zverejnila-navrh-umele-vytvorenych-koronaviru-bez-jakychkoli-rozpoznatelnych-stop-po-jejich-vyrobe","status":"publish","type":"post","link":"https:\/\/www.infokuryr.cz\/n\/2026\/05\/22\/v-roce-2008-laborator-baric-zverejnila-navrh-umele-vytvorenych-koronaviru-bez-jakychkoli-rozpoznatelnych-stop-po-jejich-vyrobe\/","title":{"rendered":"V roce 2008 laborato\u0159 Baric zve\u0159ejnila n\u00e1vrh um\u011ble vytvo\u0159en\u00fdch koronavir\u016f bez jak\u00fdchkoli rozpoznateln\u00fdch stop po jejich v\u00fdrob\u011b"},"content":{"rendered":"
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Pomoc\u00ed techniky \u201eNo See\u2019m\u201c se Baricovi a jeho koleg\u016fm poda\u0159ilo odstranit z hotov\u00e9ho genomu koronaviru v\u0161echny viditeln\u00e9 stopy sestavov\u00e1n\u00ed genomu v laborato\u0159i.<\/span><\/strong><\/p>\n

V kapitole o metod\u00e1ch publikovan\u00e9 v roce 2008 v laborato\u0159i Ralpha Barice byl kompletn\u00ed protokol pro digit\u00e1ln\u00ed konstrukci genom\u016f koronaviru otev\u0159en\u011b publikov\u00e1n v pln\u00e9m zn\u011bn\u00ed na po\u010d\u00edta\u010di, na\u010de\u017e byly tyto genomy fyzicky sestaveny do fin\u00e1ln\u00ed sekvence genomu, p\u0159i\u010dem\u017e z t\u00e9to sekvence byly odstran\u011bny v\u0161echny viditeln\u00e9 stopy genetick\u00e9 manipulace.<\/span><\/p>\n

Kapitolu s n\u00e1zvem \u201e\u00a0<\/span>Systematick\u00e9 sestavov\u00e1n\u00ed a genetick\u00e1 manipulace s genomem my\u0161\u00edho viru hepatitidy A59<\/span><\/a>\u00a0\u201c napsali Eric F. Donaldson, Amy C. Sims a Ralph S. Baric a publikovali ji v\u00a0<\/span>Methods in Molecular Biology<\/span><\/em>\u00a0, kni\u017en\u00ed s\u00e9rii nakladatelstv\u00ed Springer\/Humana Press, kter\u00e1 obsahuje laboratorn\u00ed protokoly.<\/span><\/p>\n

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St\u00e1hnout 978 1 59745 181 9 Kapitola 21, 707 KB \u2219 PDF soubor<\/span><\/a><\/strong><\/p>\n

J\u00e1drem syst\u00e9mu je technologie \u201eNo See’m\u201c.<\/span><\/p>\n

Baric tuto techniku \u200b\u200bji\u017e popsal v publikaci z roku 2006 takto:<\/span><\/p>\n

\u201eM\u00edsta bez viditelnosti lze pou\u017e\u00edt k vlo\u017een\u00ed ciz\u00edch gen\u016f do virov\u00fdch, eukaryotick\u00fdch nebo mikrobi\u00e1ln\u00edch genom\u016f \u010di vektor\u016f a z\u00e1rove\u0148 odstranit v\u0161echny stopy restrik\u010dn\u00edch m\u00edst pou\u017eit\u00fdch p\u0159i manipulaci s rekombinantn\u00ed DNA.\u201c<\/span><\/em><\/p>\n

V kapitole z roku 2008 auto\u0159i vysv\u011btluj\u00ed, jak to funguje v praxi s koronaviry:<\/span><\/p>\n

\u201eVyvinuli jsme platformu pro sestaven\u00ed DNA, kter\u00e1 vyu\u017e\u00edv\u00e1 nespecifick\u00e9, vysoce variabiln\u00ed sekven\u010dn\u00ed signatury restrik\u010dn\u00edch enzym\u016f typu IIs k sestaven\u00ed molekul\u00e1rn\u00edho klonu kmene A59 my\u0161\u00edho koronaviru hepatitidy (MHV) v pln\u00e9 d\u00e9lce. Tento p\u0159\u00edstup tak\u00e9 umo\u017e\u0148uje vlo\u017een\u00ed modifikac\u00ed do fragmentu DNA navr\u017een\u00edm primer\u016f obsahuj\u00edc\u00edch restrik\u010dn\u00ed m\u00edsto a po\u017eadovan\u00e9 mutace. P\u0159id\u00e1n\u00edm restrik\u010dn\u00edho m\u00edsta typu IIs ve spr\u00e1vn\u00e9 orientaci n\u00e1sledn\u00e9 \u0161t\u011bpen\u00ed odstran\u00ed restrik\u010dn\u00ed m\u00edsto a zanech\u00e1 lepkav\u00fd konec obsahuj\u00edc\u00ed po\u017eadovanou mutaci, p\u0159ipraven\u00fd k ligaci s druh\u00fdm fragmentem generovan\u00fdm paraleln\u011b jako jeho komplement. V t\u00e9to kapitole se budeme zab\u00fdvat detaily metody sestaven\u00ed infek\u010dn\u00edho klonu MHV v pln\u00e9 d\u00e9lce a n\u00e1sledn\u00fdm vlo\u017een\u00edm specifick\u00e9 mutace do klonu, abychom demonstrovali s\u00edlu tohoto unik\u00e1tn\u00edho p\u0159\u00edstupu \u201ene\u201c specifick\u00e9ho pro dan\u00e9 m\u00edsto, kter\u00fd demonstruje mutagenezi See’m.\u201c<\/span><\/em><\/p>\n

Rozd\u011blili genom koronaviru na sedm modul\u00e1rn\u00edch fragment\u016f cDNA.<\/span><\/p>\n

Pomoc\u00ed po\u010d\u00edta\u010dem podporovan\u00fdch primer\u016f zavedli do ka\u017ed\u00e9ho fragmentu do\u010dasn\u00e1 restrik\u010dn\u00ed m\u00edsta typu II.<\/span><\/p>\n

Tyto \u010d\u00e1sti byly po digesci zcela odstran\u011bny.<\/span><\/p>\n

V\u00fdsledn\u00e9 lepkav\u00e9 konce bez jizev byly ligov\u00e1ny dohromady za vzniku cDNA o pln\u00e9 d\u00e9lce obsahuj\u00edc\u00ed pouze p\u0159esnou sekvenci naprogramovanou v\u011bdci v po\u010d\u00edta\u010di \u2013 bez zb\u00fdvaj\u00edc\u00edch restrik\u010dn\u00edch m\u00edst, linker\u016f nebo jin\u00fdch laboratorn\u00edch marker\u016f.<\/span><\/p>\n

Bez techniky \u201eno see’m\u201c by kone\u010dn\u011b sestaven\u00fd genom \u00fadajn\u011b vykazoval zjevn\u00e9 laboratorn\u00ed stopy: dal\u0161\u00ed m\u00edsta pro restrik\u010dn\u00ed enzymy na ka\u017ed\u00e9m spojen\u00ed, linkerov\u00e9 sekvence, duplikovan\u00e9 motivy a dal\u0161\u00ed stopy um\u011bl\u00e9ho klonov\u00e1n\u00ed, kter\u00e9 by okam\u017eit\u011b odhalily, \u017ee genom byl roz\u0159ez\u00e1n a znovu sestaven v laborato\u0159i.<\/span><\/p>\n

Publikace uv\u00e1d\u00ed, \u017ee platforma je ji\u017e v provozu a \u0161iroce vyu\u017e\u00edv\u00e1na:<\/span><\/p>\n

\u201eDo klonu MHV bylo t\u00edmto p\u0159\u00edstupem vlo\u017eeno v\u00edce ne\u017e sto mutant\u016f, co\u017e daleko p\u0159evy\u0161uje po\u010det v\u0161ech ostatn\u00edch d\u0159\u00edve popsan\u00fdch reverzn\u00edch genetick\u00fdch syst\u00e9m\u016f pro koronaviry. Tato technologie byla nav\u00edc pou\u017eita k rychl\u00e9mu v\u00fdvoji \u010dinidel v reakci na nov\u011b identifikovan\u00e9, vznikaj\u00edc\u00ed koronaviry.\u201c<\/span><\/em><\/p>\n

Ji\u017e v roce 2008 publikovala Baricova laborato\u0159 p\u0159esn\u00fd molekul\u00e1rn\u00ed scaffold pot\u0159ebn\u00fd k vytvo\u0159en\u00ed po\u010d\u00edta\u010dem generovan\u00fdch, bezestop\u00fdch genom\u016f koronaviru \u2013 fin\u00e1ln\u00edch sekvenc\u00ed, kter\u00e9 by mohly b\u00fdt navr\u017eeny tak, aby vypadaly jako p\u0159irozen\u011b cirkuluj\u00edc\u00ed kmen.<\/span><\/p>\n

Mohla by zcela bezestop\u00e1 \u201eneviditeln\u00e1\u201c konstrukce genomov\u00e9ho in\u017een\u00fdrstv\u00ed, kterou v roce 2008 publikovala laborato\u0159 Ralpha Barice \u2013 schopnost digit\u00e1ln\u011b navrhovat a fyzicky sestavovat genomy koronaviru v pln\u00e9 d\u00e9lce a z\u00e1rove\u0148 zcela odstranit jak\u00fdkoli laboratorn\u00ed podpis z fin\u00e1ln\u00ed sekvence \u2013 vysv\u011btlit, pro\u010d skute\u010dn\u00fd p\u016fvod SARS-CoV-2 z\u016fst\u00e1v\u00e1 navzdory let\u016fm intenzivn\u00edho mezin\u00e1rodn\u00edho v\u00fdzkumu nejasn\u00fd?<\/span><\/p>\n

Od Jona Fleetwooda<\/span><\/em><\/strong><\/p>\n<\/div>\n<\/div>\n

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Zdroj<\/span><\/a><\/span><\/h3>\n

 <\/p>\n<\/div>\n<\/div>\n","protected":false},"excerpt":{"rendered":"

Pomoc\u00ed techniky \u201eNo See\u2019m\u201c se Baricovi a jeho koleg\u016fm poda\u0159ilo odstranit z hotov\u00e9ho genomu koronaviru…<\/p>\n","protected":false},"author":2,"featured_media":107120,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":[],"categories":[7],"tags":[345,1361,1156],"_links":{"self":[{"href":"https:\/\/www.infokuryr.cz\/n\/wp-json\/wp\/v2\/posts\/107119"}],"collection":[{"href":"https:\/\/www.infokuryr.cz\/n\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.infokuryr.cz\/n\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.infokuryr.cz\/n\/wp-json\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/www.infokuryr.cz\/n\/wp-json\/wp\/v2\/comments?post=107119"}],"version-history":[{"count":0,"href":"https:\/\/www.infokuryr.cz\/n\/wp-json\/wp\/v2\/posts\/107119\/revisions"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.infokuryr.cz\/n\/wp-json\/wp\/v2\/media\/107120"}],"wp:attachment":[{"href":"https:\/\/www.infokuryr.cz\/n\/wp-json\/wp\/v2\/media?parent=107119"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.infokuryr.cz\/n\/wp-json\/wp\/v2\/categories?post=107119"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.infokuryr.cz\/n\/wp-json\/wp\/v2\/tags?post=107119"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}