Joang ho tseba hore na mahloriso a nepahetseng ke afe?
Bakeng sa inducer IPTG (isopropyl-beta-d-thiogalactoside), ho phahamisa khatello, ho ba betere.Khatiso e nepahetseng e itšetlehile ka maemo a khethehileng a liteko le phello e lakatsehang ea induction.
Ka kakaretso, khatello ea IPTG e sebelisoa sebakeng sa 0.1-1 mmM.Ho tsepamisa maikutlo ho tlase ho ka fokotsa litlamorao tse mpe ho kholo ea lisele mme ho ka fokotsa cytotoxicity ka lebaka la ho fetella ha liprotheine tse shebiloeng.Ho tsepamisa mohopolo ho phahameng ho ka baka boima bo feteletseng ba metabolism ea lisele, ho ama kholo ea lisele le ho sebetsa hantle ha polelo.
Mokhoa oa ho tseba hore na mahloriso a nepahetseng ke afe e ka ba ho lekola boemo ba polelo ea protheine e shebiloeng ka ho etsa liteko tsa induction tsa IPTG maemong a fapaneng.Liteko tse nyane tsa setso li ka etsoa ho sebelisoa mefuta e mengata ea IPTG (mohlala, 0.1 mM, 0.5 mM, 1 mM, joalo-joalo) 'me phello ea polelo ka mekhahlelo e fapaneng e ka hlahlojoa ka ho lemoha boemo ba polelo ea protheine e lebeletsoeng (mohlala ho lemoha letheba kapa fluorescence).Ho ea ka liphetho tsa liteko, mahloriso a nang le phello e ntle ka ho fetisisa ea polelo e khethiloe e le mahloriso a nepahetseng.
Ntle le moo, o ka boela oa bua ka lingoliloeng tse amehang kapa boiphihlelo ba lilaboratori tse ling ho utloisisa mefuta ea mahloriso ea IPTG e sebelisoang hangata tlasa maemo a tšoanang a liteko, ebe o ntlafatsa le ho lokisa ho latela litlhoko tsa liteko.
Ke habohlokoa ho hlokomela hore mahloriso a nepahetseng a ka 'na a fapana ho itšetlehile ka mekhoa e fapaneng ea ho hlahisa maikutlo, liprotheine tse lebisitsoeng, le maemo a liteko, kahoo ho molemo ho ntlafatsa ho latela maemo a itseng.
Nako ea poso: Sep-28-2023