IPTG CAS: 367-93-1 Ixabiso lomvelisi
I-Isopropyl β-D-1-thiogalactopyranoside (IPTG) yi-analog synthetic ye-lactose esetyenziswa ngokuqhelekileyo kuphando lwebhayoloji ye-molekyuli kunye nezicelo ze-biotechnology.I-IPTG isetyenziselwa ukukhuthaza ukubonakaliswa kwejene kwiinkqubo zebhaktheriya, apho isebenza njenge-molecular trigger ukuqalisa ukukhutshelwa kweejene ezijoliswe kuzo.
Xa ifakwe kwi-medium yokukhula, i-IPTG ithathwa yibhaktheriya kwaye inokubophelela kwiprotheni ye-lac repressor, ukuyikhusela ekuvimbeleni umsebenzi we-lac operon.I-lac operon yiqela lezakhi zofuzo ezibandakanyekayo kwi-lactose metabolism, kwaye xa iprotheni ye-repressor isusiwe, iijini zibonakaliswa.
I-IPTG isoloko isetyenziswa ngokubambisana ne-lacUV5 i-mutant promotioner, eyinguqulelo esebenzayo yokukhuthaza i-lac.Ngokudibanisa i-IPTG yokungeniswa kunye nalo mgqugquzeli oguqukayo, abaphandi banokufikelela kumanqanaba aphezulu e-gene expression.Oku kuvumela ukuveliswa kweeprotheyini ezininzi zokuhlanjululwa okanye ezinye izicelo ezisezantsi.
Ukongeza kwi-gene expression, i-IPTG ikwasetyenziswa rhoqo kwiimvavanyo zokuhlola eziluhlaza/ezimhlophe.Kobu buchule, ijini ye-lacZ idla ngokudityaniswa kwimfuza yomdla, kwaye iibhaktheriya ezivakalisa ngempumelelo le gene yokudibanisa ziya kuvelisa i-enzyme esebenzayo ye-β-galactosidase.Xa i-IPTG yongezwa kunye ne-substrate ye-chromogenic efana ne-X-gal, ibhaktheriya ebonisa i-fusion gene ijika ibe luhlaza okwesibhakabhaka ngenxa yomsebenzi we-β-galactosidase.Oku kuvumela ukuchongwa kunye nokukhethwa kweentlobo eziphindaphindiweyo eziye zadibanisa ngempumelelo i-gene yomdla.
Ukuqaliswa kwe-gene expression: I-IPTG isetyenziswa ngokuqhelekileyo ukukhuthaza ukubonakaliswa kweejene ezijoliswe kuzo kwiinkqubo zebhaktheriya.Ilinganisa i-lactose ye-inducer yendalo kwaye ibophelela kwiprotheni ye-lac repressor, ukuyikhusela ekuvimbeleni i-lac operon.Oku kuvumela ushicilelo kunye nokubonakaliswa kofuzo olufunwayo.
Ukubonakaliswa kweprotheyini kunye nokuhlanjululwa: I-IPTG induction isoloko isetyenziselwa ukuvelisa ubuninzi beeprotheyini eziphinda ziphinde zenziwe ngeenjongo ezahlukeneyo, ezifana nezifundo ze-biochemical, imveliso yonyango, okanye uhlalutyo lwesakhiwo.Ngokusebenzisa i-vectors ye-expression efanelekileyo kunye ne-IPTG induction, abaphandi banokufikelela kumanqanaba aphezulu okuveliswa kweeprotheyini ezijoliswe kwimikhosi yebhaktheriya.
Ukuhlolwa okuluhlaza okwesibhakabhaka / okumhlophe: I-IPTG isetyenziswa rhoqo ngokudibanisa ne-lacZ gene kunye ne-chromogenic substrate, njenge-X-gal, kwii-assays zokuhlola eziluhlaza / ezimhlophe.I-lacZ gene idityaniswe ngokuqhelekileyo kwi-gene yomdla, kwaye ibhaktheriya evakalisa ngempumelelo le gene ye-fusion iya kuvelisa i-enzyme ye-β-galactosidase esebenzayo.Xa i-IPTG kunye ne-substrate ye-chromogenic yongezwa, iintlobo eziphinda ziphinde zibonise i-fusion gene zijika zibe luhlaza okwesibhakabhaka, okuvumela ukuchongwa lula kunye nokukhetha.
Uphononongo lolawulo lwemfuza: I-IPTG yokungeniswa iqhele ukusetyenziswa kuphando ukufunda ukulawulwa kofuzo kunye nee-operon, ngakumbi i-lac operon.Ngokulawula ugxininiso lwe-IPTG kunye nokubeka iliso ekubonakalisweni kwamacandelo e-lac operon, abaphandi banokuphanda iindlela zolawulo lwemfuza kunye nendima yezinto ezahlukeneyo okanye utshintsho.
Iinkqubo zokubonisa imfuza: I-IPTG licandelo elibalulekileyo kwiinkqubo ezininzi zokubonisa imfuza, njengeenkqubo ezisekelwe kumxhasi we-T7.Kwezi nkqubo, umgqugquzeli we-lac uhlala esetyenziselwa ukuqhuba intetho ye-T7 RNA polymerase, leyo, iguqulela iijene ezijoliswe kuyo phantsi kolawulo lwe-T7 yokulandelelana komgqugquzeli.I-IPTG isetyenziselwa ukuphembelela ukubonakaliswa kwe-T7 RNA polymerase, ekhokelela ekusebenziseni i-target gene expression.
| Ukuqamba | C9H18O5S |
| Isivavanyi | 99% |
| Imbonakalo | Umgubo omhlophe |
| Inombolo yeCAS. | 367-93-1 |
| Ukupakisha | Incinci kwaye ininzi |
| Beka ubomi kwishelufa | iminyaka eyi-2 |
| Ugcino | Gcina kwindawo epholileyo neyomileyo |
| Isiqinisekiso | ISO. |
