Iptg hmoovtuaj yeem siv rau qhov chaw ua tshuaj loog hauv zos, nws yog siv los ua tus neeg sawv cev sib kis rau tom qab-kev kho qhov txhab mob thiab mob plab. Molecular biological reagents feem ntau siv nyob rau hauv xiav thiab dawb qhov chaw tshuaj ntsuam thiab IPTG induced protein qhia nyob rau hauv cov kab mob. IPTG, tag nrho lub npe isopropyl- - D-thiogalactoside, CAS 367-93-1, molecular mis C9H18O5S, molecular hnyav 384.37 Daltons, belongs rau cov tshuaj molecule me me. Dawb los yog yuav luag dawb hmoov, nrog tsawg solubility nyob rau hauv dej tab sis zoo solubility nyob rau hauv cov organic solvents. Muaj cov ionic pawg hauv cov molecules, yog li lawv muaj qee qhov kev coj ua hauv dej. Nyob ruaj khov ntawm chav tsev kub, tab sis yooj yim rau decompose nyob rau hauv kub los yog muaj zog acid thiab alkali tej yam kev mob. Tsis muaj ntxhiab tsw ntxhiab, tab sis me ntsis organic compound tsw yuav raug tso tawm thaum siv. Nws yog ib qho siv ntau inducer siv los ua kom muaj protein ntau hauv cov kab mob. Hauv kev kuaj mob, IPTG tuaj yeem ua haujlwm raws li kev sojntsuam fluorescent lossis tus neeg sawv cev chromogenic rau kev kuaj xyuas cov molecules lossis cov ntaub so ntswg hauv cov qauv. Los ntawm kev sib txuas nrog lub hom phiaj molecules, IPTG tuaj yeem tsim cov teeb liab fluorescent lossis hloov xim, muab kev txhawb zog rau kev kuaj mob.
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Morphological |
Crystalline hmoov |
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Xim |
Dawb |
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Melting point |
105℃C |
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Boiling point |
350.9 ° C (kwv yees kwv yees) |
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Qhov ntom |
1.3329 (zeeg) |
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Cia tej yam kev mob |
2-8 ° C |
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solubility cawv |
soluble soluble 40 feem ntawm cov kuab tshuaj |
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Acidity coefficient (pKa) |
13.00 ± 0.70 (Tshaj tawm) |
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Soluble hauv dej |
50 mg / ml |
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Flash point |
197.8 hli |
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Solubility |
1.6g/l |
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Vapor ntom ntom |
5.21 (vs huab cua) |
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Refractive Index |
1.5060 (kwv yees) |
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|
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Iptg hmoov,Lub cev tsis muaj zog, Mp. : 114 ~ 121℃C , Kev piav qhia ntawm kev siv , Cov tshuaj molecular biology reagents, feem ntau siv xiav - kuaj dawb thiab IPTG-induced protein qhia hauv cov kab mob. , Ntim specifications, 1G, 5G, 25G, 100G, 1KG, Cia tej yam kev mob
2-8℃refrigerated, tiv thaiv los ntawm lub teeb, Hazard piav qhia, Danger code: Xi, Risk theem: R36 / 37 / 38, Kev nyab xeeb theem: S23-24 / 25-36.
IPTG feem ntau yog siv rau hauv kev sim cloning uas yuav tsum tau ua rau -galactosidase kev ua haujlwm. Nws feem ntau yog siv ua ke nrog X-Gal lossis Bluo-Gal rau xiav-kev tshuaj ntsuam dawb ntawm cov kab mob sib xyaw ua ke, uas tuaj yeem raug ntxias los ntawm Chemicalbook los qhia lac operon hauv E. coli. IPTG khi rau lacI repressor protein thiab hloov nws qhov kev sib haum xeeb los tiv thaiv qhov inhibition ntawm lacZ gene encoded los ntawm -galactosidase.
IPTG, lub npe tag nrho isopropyl- -D-thiogalactoside yog ib qho kev siv inducer dav siv rau inducing thiab regulating protein qhia. Vim nws cov qauv tshwj xeeb thiab kev ua haujlwm, IPTG feem ntau yog npaj sivIptg hmoovcov txheej txheem synthesis hauv chav kuaj.
Synthetic Route
1. Glycoside synthesis:
Thawj synthesis -D-thiogalactoside feem ntau yog tsim los ntawm cov txheej txheem ntawm glycoside synthesis, los ntawm condensing galactose nrog cov hauv paus sib xws (xws li isopropyl). Cov kauj ruam no yuav tsum tau siv cov pab pawg tiv thaiv kom tsis txhob tsim los ntawm - cov khoom lag luam hauv cov tshuaj tiv thaiv tom ntej.
2. Phosphorylation cov tshuaj tiv thaiv:
Raws li glycoside synthesis, phosphate pawg tau qhia los ntawm phosphorylation cov tshuaj tiv thaiv kom tau txais isopropyl pawg - -D-thiogalactoside-5'-phosphate ester. Cov kauj ruam no yuav tsum tau siv cov phosphates thiab coj cov anhydrides lossis acids.
3. Pab pawg tiv thaiv:
Los ntawm deprotection pab pawg neeg cov tshuaj tiv thaiv, pab pawg tiv thaiv hauv glycoside raug tshem tawm kom tau txais cov khoom lag luam phiaj xwm isopropyl- - D-thiogalactoside. Cov kauj ruam no yuav tsum tau siv cov kua qaub los yog alkali tov los pab txhawb cov tshuaj tiv thaiv.
Kev sim cov kauj ruam:
Npaj reagents thiab ntsuas:
Npaj cov suab thaj tsim nyog, cov hauv paus, pab pawg tiv thaiv, phosphates, anhydrides lossis acids, cov kuab tshuaj, thiab lwm yam reagents, nrog rau cov cuab yeej sim tsim nyog xws li stirrers, thermometers, spectrophotometers, thiab lwm yam.
Synthesis ntawm glycosides:
Tshav kub thiab do galactose nrog cov hauv paus sib xws hauv cov kuab tshuaj, ntxiv cov catalyst, thiab txhawb cov tshuaj tiv thaiv condensation. Cov kauj ruam no yuav tsum tau tswj kom nruj ntawm qhov kub thiab txias lub sij hawm los xyuas kom meej qhov kev vam meej ntawm cov tshuaj tiv thaiv.
Phosphorylation Reaction:
Cov synthesized glycosides yog rhuab thiab nplawm nrog phosphate, anhydride, los yog acid nyob rau hauv ib tug hnyav, thiab ib tug catalyst yog ntxiv los txhawb phosphorylation cov tshuaj tiv thaiv. Cov kauj ruam no yuav tsum tau tswj qhov kub thiab txias lub sij hawm stirring, thaum them nyiaj rau lub polarity thiab ntau npaum li cas ntawm cov kuab tshuaj los xyuas kom meej qhov kev vam meej ntawm cov tshuaj tiv thaiv thiab tiam ntawm cov khoom.
Pab pawg tiv thaiv:
Tshem tawm cov pab pawg tiv thaiv los ntawm cov khoom phosphorylation nyob rau hauv cov kua qaub los yog alkali tov kom tau lub hom phiaj cov khoom isopropyl- - D-thiogalactoside. Cov kauj ruam no yuav tsum tswj hwm tus nqi pH thiab qhov kub thiab txias, thaum ua tib zoo saib xyuas qhov polarity thiab ntau npaum li cas ntawm cov kuab tshuaj los xyuas kom meej qhov kev vam meej ntawm cov tshuaj tiv thaiv thiab tiam ntawm cov khoom.
Sib cais thiab purification:
Lub hom phiaj cov khoom yog cais los ntawm cov tshuaj tiv thaiv sib tov los ntawm kem chromatography, recrystallization, thiab lwm yam kev sib cais thiab purification txoj kev. Cov kauj ruam no yuav tsum tau xyuam xim rau kev khiav hauj lwm xws li kub, hnyav npaum li cas, txaus tus nqi, thiab lwm yam.
Kev txheeb xyuas thiab kev txheeb xyuas:
Kev txheeb xyuas tus qauv ntawm cov khoom lag luam yog ua los ntawm kev tshuaj ntsuam xyuas xws li nuclear magnetic resonance thiab huab hwm coj spectrometry. Cov kauj ruam no yuav tsum tau siv cov cuab yeej sib txuas, cov cuab yeej siv, thiab cov cuab yeej cuab tam kom paub meej cov qauv thiab kev huv ntawm cov khoom.
Iptg hmoovLub npe tag nrho isopropyl- - D-thiogalactoside yog ib qho kev siv inducer dav siv rau inducing thiab regulating protein qhia.
1. Protein expression induction:
IPTG plays lub luag haujlwm tseem ceeb hauv kev nthuav qhia cov protein. Nws yog ib qho siv dav siv inducer uas tuaj yeem ua kom muaj txiaj ntsig zoo rau kev qhia ntawm cov noob tshwj xeeb hauv cov kab mob, yog li tau txais lub hom phiaj xav tau protein nyob rau lub sijhawm luv luv. Nws lub luag haujlwm ntawm kev ua haujlwm yog txhawm rau khi rau lacI noob ntawm cov kab mob, inhibit qhov kev ua ntawm nws cov kev tswj hwm cov protein, yog li qhib lub lac operon ntawm cov kab mob thiab pib qhia cov noob hom phiaj. Thaum siv lactose operons raws li kev txhawb nqa rau kev qhia cov protein, cov inducers yuav tsum tau, thiab IPTG tuaj yeem ua raws li lactose analogue los txhawb kev qhia ntawm galactosidase hauv Escherichia coli. Nws tsis tuaj yeem siv los ntawm cov hlwb kom ua tiav cov lus qhia. IPTG khi rau lac|cov khoom, ua rau muaj kev hloov pauv ntawm lacO, yog li ua kom muaj kev hloov pauv. Txoj cai tswjfwm tsis tu ncua no tau dhau los ua ib qho kev siv feem ntau hauv kev tsim kho E. coli kev qhia system vectors.
2. Gene expression regulation:
IPTG plays lub luag haujlwm tseem ceeb hauv kev tswj cov noob caj noob ces. Raws li ib qho tseem ceeb kev sim reagent, nws plays lub luag hauj lwm tseem ceeb nyob rau hauv cov kev cai ntawm noob caj noob ces thiab protein overexpression thwmsim. 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 cai tswjfwm no yog raws li kev khi ntawm IPTG rau lac|Cov khoom lag luam hauv lactose operons, hloov pauv lawv qhov kev sib haum xeeb, yog li tawm hauv lacO thiab ua kom muaj kev hloov pauv ntxiv. Txoj cai tswjfwm tsis raug cai no ua rau IPTG muaj qhov tseem ceeb hauv kev tshawb fawb xws li kev ua haujlwm ntawm noob, kev sib cuam tshuam ntawm cov protein, thiab kev tshuaj xyuas tshuaj.
3. Protein crystallization:
Hauv kev sim protein crystallization, IPTG yog siv los ua tus inducer los txhawb cov protein crystallization. Nws cov txheej txheem ntawm kev ua haujlwm yog hloov pauv cov qauv ntawm cov protein los ntawm kev khi rau thaj tsam hydrophobic hauv cov protein, yog li txhawb kev sib sau ua ke thiab crystallization ntawm cov protein molecules. Cov txheej txheem sib sau ua ke no thiab crystallization yog thim rov qab, yog li cov protein crystallization tuaj yeem raug ntxias los ntawm kev ntxiv IPTG, los yog cov muaju protein tuaj yeem yaj los ntawm kev tshem tawm IPTG.
Hauv kev sim protein crystallization, IPTG feem ntau yog ntxiv rau cov kua protein raws li qhov kawg ntawm 1mM. Qhov tsis tshua muaj siab ntawm IPTG tuaj yeem zam dhau qhov induction ntawm cov protein, yog li ua tiav cov teebmeem crystallization zoo dua. Nyob rau tib lub sijhawm, IPTG tuaj yeem ua haujlwm raws li molecular chaperone los pab hauv qhov tseeb folding thiab sib sau ua ke ntawm cov proteins, yog li tau txais cov khoom siv muaj protein ntau dua.
Nws yuav tsum raug sau tseg tias IPTG tsis txhawb nqa cov protein crystallization hauv txhua kis. Qee cov proteins tsis rhiab rau IPTG, lossis vim lawv cov qauv tsim thiab cov khoom uas tsis haum rau crystallization, kev sim validation thiab optimization yog xav tau rau cov proteins sib txawv los txiav txim qhov zoo crystallization tej yam kev mob thiab txoj kev.
Kev phiv tshuaj
Cytotoxicity
Toxicity rau cov cell prokaryotic
Txawm hais tias IPTG feem ntau yog siv los ntxias cov noob qhia hauv cov hlwb prokaryotic, qhov siab ntawm IPTG yuav muaj cov tshuaj lom ntawm cov hlwb prokaryotic. Kev tshawb fawb tau pom tias thaum IPTG concentration siab dhau lawm, nws tuaj yeem cuam tshuam cov txheej txheem metabolic hauv cov hlwb. Piv txwv li, nws tuaj yeem cuam tshuam rau lub zog metabolism ntawm cov hlwb, cuam tshuam rau txoj hauv kev xws li glycolysis thiab tricarboxylic acid voj voog, ua rau txo qis hauv cov cellular ATP ntau lawm thiab cuam tshuam rau kev loj hlob ntawm tes thiab kev loj hlob. Tsis tas li ntawd, cov ntsiab lus siab ntawm IPTG kuj tseem tuaj yeem ua rau kev ncaj ncees ntawm cov cell membrane, ua rau nws cov permeability, ua rau kom cov tshuaj intracellular, thiab tso cai rau cov khoom tsis zoo los ntawm ib puag ncig extracellular nkag mus rau hauv lub cell, yog li ua rau cell tuag. Cov kev sim tau pom tias thaum Escherichia coli yog kab lis kev cai hauv nruab nrab uas muaj qhov siab ntawm IPTG, qhov kev loj hlob ntawm cov kab mob ua rau qeeb qeeb thiab tus naj npawb ntawm cov kab mob siv tau maj mam txo nrog qhov txuas ntxiv ntawm lub sijhawm cog qoob loo. Qhov no qhia tau hais tias qhov siab ntawm IPTG muaj qee yam inhibitory thiab tua cov nyhuv ntawm Escherichia coli.
Toxicity rau eukaryotic hlwb
Ntxiv nrog rau cov hlwb prokaryotic, IPTG kuj tseem tuaj yeem ua rau cov hlwb eukaryotic. Muaj qee qhov sib txawv ntawm cov qauv thiab kev ua haujlwm ntawm cov hlwb eukaryotic thiab cov hlwb prokaryotic, tab sis IPTG tseem tuaj yeem cuam tshuam rau lub cev kev ua haujlwm ntawm eukaryotic hlwb los ntawm ntau txoj hauv kev. Ntawm qhov tod tes, IPTG tuaj yeem cuam tshuam nrog cov teeb liab hloov txoj hauv kev hauv eukaryotic hlwb. Cov teeb liab hloov pauv hauv cov hlwb ua lub luag haujlwm tseem ceeb hauv cov txheej txheem xws li kev loj hlob, kev sib txawv, thiab apoptosis. IPTG tuaj yeem hloov qhov kev siv zog thiab kev taw qhia ntawm kev hloov pauv ntawm cov teeb liab los ntawm kev cuam tshuam nrog qee cov cim qhia hauv cov hlwb, yog li cuam tshuam rau kev ua haujlwm ntawm lub cev ntawm cov hlwb.
Toxicity rau eukaryotic hlwb
Ntawm qhov tod tes, IPTG tuaj yeem ua rau oxidative kev ntxhov siab hauv cov hlwb eukaryotic. Oxidative stress yog hais txog qhov tsis sib xws ntawm cov oxidation intracellular oxidation thiab antioxidant kev ua, ua rau muaj kev nce hauv cov pa oxygen reactive (ROS) ntau lawm. ROS ntau dhau tuaj yeem tua cov biomolecules xws li DNA, proteins, thiab lipids hauv cell, ua rau cellular puas thiab ua haujlwm tsis zoo. Kev tshawb fawb tau pom tias nyob rau hauv tej yam kev mob, IPTG kev kho mob ntawm eukaryotic hlwb ho nce intracellular ROS theem, nrog rau ib tug txo nyob rau hauv cell viability thiab nce apoptosis tus nqi. Qhov no qhia tias IPTG tuaj yeem ua rau oxidative kev nyuaj siab teb thiab ua rau muaj toxicity ntawm eukaryotic hlwb.
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