IPTGyog ib qho kev sib xyaw tsis zoo nrog ib pawg isopropyl thiab galactose. Nws cov mis molecular yog C9H18O5S, nrog rau qhov hnyav molecular ntawm 238.30. IPTG yog soluble hauv dej thiab muaj kev ruaj ntseg siab. Hauv biology, IPTG feem ntau yog siv los ua inducer thiab tuaj yeem ua rau - Cov haujlwm ntawm galactosidase. IPTG (isopropyl) - D-thiogalactoside yog ib qho kev sim tshuaj reagent uas siv dav hauv kev lag luam ntawm molecular biology thiab genetic engineering. Nws yog ib qho kev sib xyaw ua ke nrog cov qauv zoo ib yam li cov lactose, tab sis muaj cov tshuaj sib txawv.
(Cov khoom txuas 1:https://www.bloomtechz.com/synthetic-chemical/api-researching-only/iptg-reagent-cas-367-93-1.html )
(Cov khoom txuas 1:https://www.bloomtechz.com/synthetic-chemical/api-researching-only/iptg-powder-cas-367-93-1.html )
1. Molecular qauv:
Cov tshuaj formula ntawm IPTG yog C9H18O5S, CAS 367-93-1, thiab tus txheeb ze molecular hnyav yog 238.30 g / mol. Nws cov qauv yog txuas nrog ib pawg isopropyl rau - C1 txoj hauj lwm ntawm D-thiogalactoside tsim ib daim ntawv cog lus ester. Cov qauv no ua rau IPTG simulate cov nyhuv induction ntawm lactose ntawm lactase.
Nws cov qauv molecular feem ntau muaj cov hauv qab no:
Qab zib ib feem: Cov piam thaj ib feem ntawm IPTG yog - D-galactose, zoo ib yam li thiogalactose, kuj tau lees paub los ntawm galactosidase ntawm Escherichia coli.
Thiogalactosyl moiety: Tsis zoo li cov galactose zoo tib yam, galactosyl moiety ntawm IPTG yog hloov los ntawm sulfur atoms, uas tso cai rau IPTG hloov kho los ntawm E. coli - Galactosidase paub thiab ua haujlwm raws li nws cov substrate.
Isopropyl pawg: Lwm qhov ntawm IPTG yog pawg isopropyl, uas txo cov solubility ntawm IPTG hauv dej thiab pab txhawb nws cov permeation hauv cell kab lis kev cai.
Nyob rau hauv cov nqe lus ntawm cov qauv molecular, lub substrate ntawm IPTG thiab galactosidase - D-galactoside (G1P) zoo ib yam, tshwj tsis yog hais tias ib tug sulfur atom ntxiv rau galactoside feem. Thaum IPTG raug coj los ntawm cov hlwb, nws tuaj yeem ua raws li - Lub substrate ntawm galactosidase, uas nkag mus rau hauv thiogalactose thiab isopropyl pawg nyob rau hauv qhov kev txiav txim ntawm enzyme - - D-glucose. Lub zog tso tawm los ntawm cov txheej txheem no tuaj yeem siv los tsim cov proteins txawv teb chaws.
Ntxiv rau nws cov qauv molecular cov yam ntxwv, IPTG kuj muaj ntau qhov zoo uas ua rau nws siv ntau inducer. Ua ntej, nws cov dej solubility kuj zoo thiab tuaj yeem yooj yim ntxiv rau cov kab lis kev cai nruab nrab. Qhov thib ob, nws cov nyhuv inducing ntawm Escherichia coli yog qhov me me thiab tsis ua rau lub zog ntau dhau ntawm cov hlwb, uas muaj txiaj ntsig zoo rau kev ncua lub neej ntawm cov hlwb. Tsis tas li ntawd, kev siv thiab kev siv ntawm IPTG hauv cov hlwb yog qhov nrawm heev, uas tuaj yeem ua rau cov noob qhia tau raws sijhawm.
2. Solubility:
IPTG yog cov xim tsis muaj xim crystalline nrog zoo solubility hauv dej. Nws tuaj yeem yaj sai sai ntawm chav tsev kub thiab tsim cov tshuaj pob tshab. Tsis tas li ntawd, IPTG kuj yog soluble hauv qee cov kuab tshuaj organic, xws li methanol, ethanol, thiab dimethyl sulfoxide.
3. Kev ruaj ntseg:
IPTG yog qhov ruaj khov nyob rau hauv cov kev sim ua raws li cov qauv kev sim thiab tsis yooj yim rau decomposition lossis degradation. Nws tuaj yeem khaws cia rau lub sijhawm ntev yam tsis poob nws txoj haujlwm. Txawm li cas los xij, nyob rau hauv qhov kub thiab txias los yog acidic, IPTG tuaj yeem raug cov tshuaj tiv thaiv hydrolysis, ua rau nws poob nws lub peev xwm los ntxias lactase.
4. Inducing lactase:
IPTG yog ib qho zoo inducer ntawm lactase. Nyob rau hauv feem ntau Escherichia coli, lactase yog ib qho tseem ceeb metabolic enzyme siv los zom lactose rau hauv qabzib thiab galactose. IPTG muaj cov qauv zoo sib xws rau lactose thiab tuaj yeem khi rau qhov chaw induction ntawm lactase thiab qhib nws cov ntawv sau. Qhov no ua rau IPTG yog ib qho cuab yeej tseem ceeb rau kev kawm txog noob caj noob ces txoj cai thiab kev qhia cov protein.
Hauv cov kab mob, lactose operons yog cov txheej txheem tseem ceeb uas tuaj yeem tswj cov kab mob metabolism ntawm lactose. Thaum cov piam thaj tsis muaj cov kab mob hauv cov kab mob, lactose operons raug ntxias los tsim cov enzymes uas muaj peev xwm rhuav tshem lactose.
Kev sib xyaw ntawm lactose operons: Lub lactose operon yog tsim los ntawm peb lub noob, xws li lacZ, lacY, thiab lacA. Ntawm lawv, lacZ encoding - Galactosidase, lacY encodes permeability protein, lacA encodes acetyltransferase. Peb cov noob no ua haujlwm ua ke los pab cov kab mob kom siv lactose.
Cov txheej txheem ntawm kev ua ntawm IPTG: Thaum IPTG tshwm sim, nws tuaj yeem cuam tshuam nrog - Kev khi ntawm galactosidase txhim kho cov haujlwm enzyme. Qhov kev khi no tau ua tiav los ntawm kev sib cuam tshuam ntawm pawg galactose hauv IPTG molecule thiab - Kev khi ntawm qhov chaw nquag ntawm galactosidase tiav. Qhov kev khi no ua rau muaj kev ua haujlwm ntawm enzyme, yog li txhawb kev tawg ntawm lactose.
Cov txheej txheem induction: Hauv ib puag ncig uas tsis muaj piam thaj, lacY thiab lacA cov noob tau ua ke, tab sis tus nqi ntawm cov synthesis kuj tsawg. Thaum IPTG tshwm sim, nws tuaj yeem cuam tshuam nrog - Kev khi ntawm galactosidase txhim kho cov haujlwm enzyme. Qhov kev khi no txhawb nqa kev hloov pauv ntawm lacY thiab lacA cov noob, tso cai rau cov kab mob los ua kom muaj cov protein ntau permeability thiab acetyltransferases. Cov enzymes no tuaj yeem txhawb cov kab mob metabolism ntawm lactose.
Cov yam ntxwv cuam tshuam: Qhov concentration ntawm IPTG yuav cuam tshuam rau cov nyhuv induction. Tsawg concentration ntawm IPTG tuaj yeem txhawb nqa - Lub synthesis ntawm galactosidase, tab sis siab ntau ntawm IPTG tuaj yeem muaj cov tshuaj lom rau cov hlwb. Tsis tas li ntawd, cov nyhuv induction ntawm IPTG kuj cuam tshuam los ntawm cov yam ntxwv xws li kub, pH tus nqi, thiab lub sijhawm cog qoob loo.
5. Tsis muaj toxicity:
Piv rau lactose, qhov metabolic tus nqi ntawm IPTG hauv cov hlwb qeeb qeeb, yog li nws muaj kev cuam tshuam tsawg rau kev loj hlob ntawm tes thiab cov metabolism. Qhov no ua rau IPTG feem ntau siv inducer hauv chav kuaj los tswj kev qhia ntawm cov hom phiaj.
6. Daim ntawv thov:
IPTG feem ntau yog siv rau hauv cov hauv qab no:
-Protein qhia: Los ntawm kev siv IPTG inducers, cov txiaj ntsig ntawm lub hom phiaj cov protein nyob rau hauv lub recombinant protein qhia system tuaj yeem tswj tau. Nws tuaj yeem siv los kawm txog cov txheej txheem lom neeg xws li kev ua haujlwm ntawm cov protein, kev sib cuam tshuam, thiab kev xa cov teeb liab. Piv txwv li, los ntawm kev sim cov protein crystallization, cov qauv thiab kev ua haujlwm ntawm cov protein tuaj yeem kawm tau; Los ntawm kev sim cov protein sib cuam tshuam, kev sib cuam tshuam ntawm cov protein tuaj yeem kawm tau; Los ntawm cov teeb liab transduction thwmsim, lub luag hauj lwm ntawm cov proteins nyob rau hauv lub teeb liab transduction yuav kawm tau.
-Kev tshawb fawb txog noob caj noob ces: IPTG tuaj yeem simulate lactose induction mechanism hauv cov hlwb, yog li kawm txog kev tswj cov noob caj noob ces thiab cov teeb liab hloov txoj hauv kev. Nyob rau hauv gene qhia kev cai thwmsim, IPTG pab raws li ib tug inducer thiab yuav khi rau lac|cov khoom hauv lactose operons, inducing conformational hloov thiab ua rau lac|cov khoom kom tawm ntawm qhov chaw khi ntawm tus neeg txhawb nqa, yog li ua kom cov ntaub ntawv hloov pauv. Txoj cai tswjfwm kev hloov pauv tsis tu ncua no ua rau IPTG ua lub luag haujlwm tseem ceeb hauv kev tswj hwm noob caj noob ces. Los ntawm kev tswj hwm lub sijhawm ntxiv thiab qhov siab ntawm IPTG, kev tswj hwm ntawm lub hom phiaj qhia cov protein tuaj yeem ua tiav.
-Txoj kev tshawb fawb txog kev hloov pauv: IPTG tuaj yeem siv los kawm txog kev sib cuam tshuam ntawm cov txheej txheem hloov pauv thiab lawv cov hom phiaj, nrog rau cov txheej txheem tswj hwm kev ua haujlwm ntawm cov txheej txheem hloov pauv. Los ntawm kev sib txuas nrog lub replicator ntawm lactase, IPTG tuaj yeem simulate cov txheej txheem tswj hwm ntawm lactose ntawm lactase, yog li tswj cov noob qhia. Cov txheej txheem induction no tuaj yeem siv rau kev tshawb fawb txog kev hloov pauv los tshawb nrhiav lub luag haujlwm ntawm kev hloov pauv ntawm cov noob caj noob ces. Piv txwv li, ib qho kev qhia vector uas muaj lub hom phiaj kev hloov pauv tuaj yeem tsim thiab koom ua ke nrog cov neeg txhawb nqa tsim nyog thiab kev tswj hwm cov ntsiab lus rau hauv cov lus qhia vector, ua raws li qhov sib ntxiv ntawm IPTG los txhawb qhov kev qhia ntawm lub hom phiaj transcription.
IPTG yog ib qho kev siv tshuaj reagent nrog zoo solubility thiab stability. Nws tuaj yeem cuam tshuam qhov kev qhia ntawm lactase thiab tau siv dav hauv cov haujlwm ntawm molecular biology thiab genetic engineering. Los ntawm kev siv IPTG, cov kws tshawb fawb tuaj yeem tshawb nrhiav cov teeb meem lom neeg tseem ceeb xws li cov txheej txheem tswj cov noob caj noob ces, kev qhia cov protein, thiab cov txiaj ntsig kev hloov pauv.