Dithiothreitol (DTT) ke ntho e atisang ho sebelisoa e fokotsang, e tsejoang hape ka hore ke tlatsetso e ncha e tala.Ke motsoako o monyane oa limolek'hule o nang le lihlopha tse peli tsa mercaptan (-SH).Ka lebaka la ho fokotsa thepa le botsitso, DTT e sebelisoa haholo litekong tsa biochemistry le molecular biology.
Karolo e ka sehloohong ea DTT ke ho fokotsa litlamo tsa disulfide liprotheine le li-biomolecule tse ling.Tlamahano ea disulfide ke karolo ea bohlokoa ea ho phutha ha protheine le botsitso, empa tlas'a maemo a itseng a liteko, joalo ka tlhahlobo e fokolang ea SDS-PAGE, recombination ea protheine le ho phutha, hoa hlokahala ho fokotsa tlamo ea disulfide ho lihlopha tse peli tsa thiol ho manolla sebopeho sa sebaka sa libaka. protheine.DTT e ka itšoara ka litlamo tsa disulfide ho li fokotsa ho lihlopha tsa mercaptan, kahoo li bula sebopeho sa sebaka sa protheine le ho etsa hore ho be bonolo ho hlahloba le ho laola.
DTT e ka boela ea sebelisoa ho sireletsa ts'ebetso ea enzyme le botsitso.Lits'ebetsong tse ling tsa enzyme-catalyzed, ts'ebetso ea enzyme e ka fokotsoa ke oxidant.DTT e ka sebetsana le li-oxidants ho li fokotsa ho lintho tse se nang kotsi, kahoo li sireletsa mosebetsi le botsitso ba enzyme.
Ha ho bapisoa le mekhoa ea ho fokotsa mekhoa e tloaelehileng e kang β-mercaptoethanol (β-ME), DTT e nkoa e le mokhoa o sireletsehileng le o tsitsitseng oa ho fokotsa.Ha e tsitsitse feela ka tharollo ea metsi, empa hape e boloka thepa ea eona ea ho fokotsa tlas'a mocheso o phahameng le maemo a acid-base.
Tšebeliso ea DTT e batla e le bonolo.Ka kakaretso, DTT e qhaloa ka buffer e loketseng ebe e kenngoa tsamaisong ea liteko.Boikoetliso bo nepahetseng ba DTT bo hloka ho khethoa ho latela teko e khethehileng, 'me ka kakaretso e sebelisoa ho tloha 0.1-1mM.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.
Ka kakaretso, DTT ke sesebelisoa se fokotsang se sebelisoang hangata se ka sebelisoang ho fokotsa litlamo tsa disulfide liprotheine le li-biomolecules tse ling le ho sireletsa mosebetsi oa enzyme le botsitso.E 'nile ea sebelisoa haholo litekong tsa biochemistry le molecular biology.
Nako ea poso: Sep-28-2023