IPTG CAS:367-93-1 Intengo Yomkhiqizi
I-Isopropyl β-D-1-thiogalactopyranoside (IPTG) iyi-analog yokwenziwa ye-lactose evame ukusetshenziswa ocwaningweni lwebhayoloji yamangqamuzana nasekusetshenzisweni kwe-biotechnology.I-IPTG isetshenziselwa kakhulu ukunxenxa ukuvezwa kofuzo ezinhlelweni zebhaktheriya, lapho isebenza khona njengesibasi samangqamuzana ukuqalisa ukulotshwa kofuzo oluqondiwe.
Uma ingezwe endaweni yokukhula, i-IPTG ithathwa yibhaktheriya futhi ingakwazi ukubopha iphrotheni ye-lac repressor, ukuyivimbela ekuvimbeleni umsebenzi we-lac operon.I-lac operon iyiqoqo lezakhi zofuzo ezihilelekile ku-lactose metabolism, futhi lapho iphrotheni ye-repressor isusiwe, izakhi zofuzo ziyavezwa.
I-IPTG ivamise ukusetshenziswa ngokuhambisana nomgqugquzeli we-lacUV5 oguquguqukayo, okuyinguqulo esebenzayo yomgqugquzeli we-lac.Ngokuhlanganisa ukungeniswa kwe-IPTG nalo mgqugquzeli oguquguqukayo, abacwaningi bangafinyelela amazinga aphezulu enkulumo yofuzo.Lokhu kuvumela ukukhiqizwa kwenani elikhulu lamaprotheni ukuze ahlanzwe noma ezinye izinhlelo zokusebenza ezansi nomfula.
Ngokungeziwe ekukhulumeni kofuzo, i-IPTG iphinde isetshenziswe njalo ekuhlolweni kokuhlola okuluhlaza okwesibhakabhaka/okumhlophe.Kule nqubo, isakhi sofuzo se-lacZ sivamise ukuhlanganiswa nofuzo oluthakaselwayo, futhi amagciwane aveza ngempumelelo lolu fuzo oluxubile azokhiqiza i-enzyme ye-β-galactosidase esebenzayo.Uma i-IPTG yengezwa kanye ne-substrate ye-chromogenic efana ne-X-gal, amagciwane aveza isakhi sofuzo esihlanganisiwe aphenduka abe luhlaza okwesibhakabhaka ngenxa yomsebenzi we-β-galactosidase.Lokhu kuvumela ukukhonjwa nokukhethwa kwezinhlobo ezihlanganisiwe ezihlanganise ngempumelelo ufuzo lwentshisekelo.
Ukwethulwa kwesisho sofuzo: I-IPTG ijwayele ukusetshenziselwa ukunxenxa ukuvezwa kofuzo oluqondiwe ezinhlelweni zebhaktheriya.Ilinganisa i-lactose ye-inducer yemvelo futhi ibophezela kuphrotheni ye-lac repressor, ivimbele ukuthi ingavimbeli i-lac operon.Lokhu kuvumela ukulotshwa nokuvezwa kofuzo olufunwayo.
Ukuchazwa kwamaprotheni kanye nokuhlanzwa: Ukungeniswa kwe-IPTG kuvame ukusetshenziselwa ukukhiqiza inani elikhulu lamaprotheni ahlanganisiwe ngezinjongo ezihlukahlukene, njengezifundo ze-biochemical, ukukhiqizwa kokwelapha, noma ukuhlaziywa kwesakhiwo.Ngokusebenzisa ama-vector okubonisa afanelekile kanye nokungeniswa kwe-IPTG, abacwaningi bangafinyelela amazinga aphezulu okukhiqizwa kwamaprotheni okuqondiwe kuma-bacterial host.
Ukuhlola okuluhlaza okwesibhakabhaka/okumhlophe: I-IPTG ivamise ukusetshenziswa ngokuhlanganiswa ne-lacZ gene kanye ne-chromogenic substrate, efana ne-X-gal, kuma-assays okuhlola okuluhlaza/okumhlophe.Isakhi sofuzo se-lacZ sivamise ukuhlanganisa isakhi sofuzo esithakaselwayo, futhi amagciwane aveza ngempumelelo lolu fuzo oluhlanganisayo azokhiqiza i-enzyme ye-β-galactosidase esebenzayo.Lapho i-IPTG ne-substrate ye-chromogenic yengezwa, izinhlobo eziphindayo eziveza isakhi sofuzo esixubile ziba luhlaza okwesibhakabhaka, okuvumela ukuhlonza nokukhetha kalula.
Ucwaningo lokulawulwa kofuzo: Ukungeniswa kwe-IPTG kuvame ukusetshenziswa ocwaningweni ukutadisha ukulawulwa kofuzo nama-operon, ikakhulukazi i-lac operon.Ngokukhohlisa ukugxila kwe-IPTG nokuqapha ukuvezwa kwezingxenye ze-lac operon, abacwaningi bangaphenya izindlela zokulawulwa kofuzo kanye nendima yezinto ezihlukahlukene noma ukuguqulwa.
Amasistimu wokuveza izakhi zofuzo: I-IPTG iyingxenye ebalulekile ezinhlelweni zokuchazwa kwezakhi zofuzo ezimbalwa, njengezinhlelo ezisekelwe kumgqugquzeli we-T7.Kulezi zinhlelo, umgqugquzeli we-lac uvame ukusetshenziselwa ukushayela inkulumo ye-T7 RNA polymerase, yona, ebhala izakhi zofuzo eziqondiwe ngaphansi kokulawula ukulandelana komgqugquzeli we-T7.I-IPTG isetshenziselwa ukunxenxa ukuvezwa kwe-T7 RNA polymerase, okuholela ekusebenziseni isisho sofuzo oluqondiwe.
| Ukwakheka | I-C9H18O5S |
| Isivivinyo | 99% |
| Ukubukeka | Impushana emhlophe |
| Inombolo ye-CAS. | 367-93-1 |
| Ukupakisha | Ezincane nezinkulu |
| I-Shelf Life | iminyaka engu-2 |
| Isitoreji | Gcina endaweni epholile neyomile |
| Isitifiketi | ISO. |
