Isopropylthio-beta-galactoside

Isopropylthio-beta-galactoside - Names and Identifiers

Name	Isopropyl-beta-D-thiogalactopyranoside
Synonyms	IPTG YPD BROTH E IPTG, HEMIDIOXANE ADDUCT Isopropyl thiogalactoside Isopropylthio-beta-galactoside isopropyl-thiogalactopyranoside Isopropyl-beta-D-thiogalactoside Isopropyl-β-D-thiogalactopyranoside Isopropylthio-beta-Thiogalactosidase Isopropylthio-B-thiogalactopyranoside Isopropyl β-D-1 Thiogalactopyranoside Isopropyl-beta-D-thiogalactopyranoside 1-ISOPROPYL-BETA-D-THIOGALACTOPYRANOSIDE POLYETYLENE GLYCOL 6000 MOL. BIOLOGY GRA 1-methylethyl 1-thio-D-galactopyranoside IPTG (Isopropyl-β-D-thiogalatopyranoside) Isopropyl-1-thio-beta-D-galactopyranoside propan-2-yl 1-thio-beta-D-galactopyranoside 1-methylethyl1-thio-.beta.-D-Galactopyranoside Isopropyl-beta-D-thiogalactopyranoside, Dioxane Free Isopropyl 2,3-O-isopropylidene-4-O-benzoyl-β-L-thiorhamnopyranoside
CAS	367-93-1
EINECS	206-703-0
InChI	InChI=1/C9H18O5S/c1-4(2)15-9-8(13)7(12)6(11)5(3-10)14-9/h4-13H,3H2,1-2H3/t5-,6+,7+,8-,9?/m1/s1
InChIKey	BPHPUYQFMNQIOC-NXRLNHOXSA-N

Isopropylthio-beta-galactoside - Physico-chemical Properties

Molecular Formula	C9H18O5S
Molar Mass	238.3
Density	1.3329 (rough estimate)
Melting Point	105 °C
Boling Point	350.9°C (rough estimate)
Specific Rotation(α)	-31 º (c=1, water)
Flash Point	219°C
Water Solubility	soluble
Solubility	Soluble in water, and methanol
Vapor Presure	1.58E-09mmHg at 25°C
Appearance	White powder
Color	White
Merck	14,5082
BRN	4631
pKa	13.00±0.70(Predicted)
Storage Condition	2-8°C
Stability	Stable. Incompatible with strong oxidizing agents.
Sensitive	`sensitive` to humidity and heat
Refractive Index	1.5060 (estimate)
MDL	MFCD00063273
Physical and Chemical Properties	Specific rotation -31 ° (c = 1, water) water-soluble melting point 105 °c

Isopropylthio-beta-galactoside - Risk and Safety

Risk Codes	R19 - May form explosive peroxides R40 - Limited evidence of a carcinogenic effect R66 - Repeated exposure may cause skin dryness or cracking R36/37/38 - Irritating to eyes, respiratory system and skin.
Safety Description	S23 - Do not breathe vapour. S24/25 - Avoid contact with skin and eyes. S36/37 - Wear suitable protective clothing and gloves. S22 - Do not breathe dust. S36/37/39 - Wear suitable protective clothing, gloves and eye/face protection. S27 - Take off immediately all contaminated clothing. S26 - In case of contact with eyes, rinse immediately with plenty of water and seek medical advice.
WGK Germany	3
FLUKA BRAND F CODES	10
TSCA	Yes
HS Code	29389090

Isopropylthio-beta-galactoside - Reference

Reference

1. Huang Cong, Li Xiao, Xu Chaoqun, etc. Metabolic flux of β-mannanase produced by genetically engineered Escherichia coli BL21 [J]. China Brewing, 2019, 38(02):48-52.
2. Zhao Chengcheng, Sun Changpo, Chang Xiaojiao, etc. Construction of E. Coli cell lysis system and its application in expression of mycotoxin degrading enzyme [J]. Chinese Journal of Biological Engineering, 2019, 039(004):69-77.
3. Peng Yan, Lu Chao, Qi Zhenqiang, etc. Preparation of Helicobacter pylori pilin FLaA IgY and its antibacterial activity in vitro [J]. World Chinese Journal of Gastroenterology, 2014, 29 (14):1936-442.
4. Smectite Ali abdounar, guxinorong, Ma Ji, Ma Xiaoli, Liu Xiaoning. Preparation and identification of mouse polyclonal antibody against type Ⅳ chitinase (HaCHT4) of cotton bollworm [J]. Journal of Cellular and Molecular Immunology, 2018,34(09):845-849.
5. Nianyuan, Deng Yuanyuan, Wei Zhencheng, Zhang Yan, Tang Xiaojun, Li Ping, Zhou Pengfei, Liu Guang, Zhang famous. Expression and enzymatic characteristics of recombinant wheat resting thiol oxidase and its effect on bread quality [J]. Chinese Journal of bioengineering, 2021,37(02):593-603.
6. [IF = 2.1] Fei Wu et al."Recombinant human histidine triad nucleotide-binding protein 1 attenuates liver fibrosis induced by carbon tetrachloride in rats."Mol Med Rep. 2013 Oct;8(4):1023-1028
7. [IF=6.119] Peng Jin et al."Efficient bioconversion of high-concentration D-fructose into D-mannose by a novel N-acyl-D-glucosamine 2-epimerase from Thermobifida halotolerans."Catal Sci Technol. 2021 Mar;11(5):1922-1930

Isopropylthio-beta-galactoside - Introduction

IPTG is an activity-inducing substance of β-galactosidase. Based on this characteristic, when the vector DNA of pUC series (or other vector DNA with lacZ gene) is transformed with lacZ deletion cells as the host, or when the vector DNA of M13 phage is transfected, if X-gal and IPTG are added to the plate medium, due to the α-complementarity of β-galactosidase, the gene recombinant can be easily selected according to whether white colonies (or plaques) appear. In addition, it can also be used as an expression inducer for expression vectors with promoters such as lac or tac. Soluble in water, methanol, ethanol, soluble in acetone, chloroform, insoluble in ether. It is an inducer of β-galactosidase and β-galactosidase. It is not hydrolyzed by β-galactoside. It is a substrate solution of thiogalactosyltransferase. Formulated: IPTG is dissolved in water, and then sterilized to prepare a storage solution (0 · 1M). The final IPTG concentration in the indicator plate should be 0 · 2mM.

Last Update：2022-10-16 17:24:55

Isopropylthio-beta-galactoside - Reference Information

EPA chemical information	Information provided by: ofmpub.epa.gov (external link)
overview	isopropyl thiogalactoside (IPTG) is a very strong inducer, which is not metabolized by bacteria and is very stable, so it is widely used in laboratories. IPTG is often used in cloning experiments requiring induction of β-galactosidase activity. It is often used in combination with X-Gal or Bluo-Gal for blue-white screening of recombinant bacterial colonies, which can induce the expression of lac operon in Escherichia coli. IPTG binds to lacI repressor protein and changes its conformation to play a role in preventing the inhibition of the β-galactosidase encoding gene lacZ.
principle of induction	The lactose operon (element) of E.coli contains three structural genes Z, Y and A, which encode galactosidase, peroxidase and acetyltransferase respectively. In addition, there is a manipulation sequence O, a starting sequence P and a regulatory gene I. The I gene encodes a repressor protein that binds to the O sequence, causing the operon (element) to be repressed and turned off. There is also a catabolite gene activator protein (CAP) binding site upstream of the priming sequence P. The regulatory region of lac operon is composed of P sequence, O sequence and CAP binding site. The coding genes of the three enzymes are regulated by the same regulatory region to realize the coordinated expression of gene products. When there is no lactose, the lac operon (element) is in a repressive state. At this time, the Lac repressor protein expressed by the I sequence under the manipulation of the PI start sequence binds to the O sequence, preventing the RNA polymerase from binding to the P sequence and inhibiting the transcription start. When lactose is present, the lac operon (element) can be induced. In this operon (meta) system, the real inducer is not lactose itself. Lactose enters the cell and is catalyzed by β-galactosidase and converted into isolactose. As an inducer, the latter molecule binds to the repressor protein to change the conformation of the protein, leading to the dissociation and transcription of the repressor protein from the O sequence. The effect of isopropyl thiogalactoside (IPTG) is the same as that of isolactose. It is a very strong inducer, which is not metabolized by bacteria and is very stable, so it is widely used in laboratories.
advantages of IPTG	1. molecules that can induce enzyme synthesis but are not decomposed are called comfort inducers. Although lactose can induce the synthesis of enzymes, it decomposes with it, resulting in many complex kinetic problems. Therefore, people often use comfort inducers to carry out various experiments. 2.IPTG is not metabolized by bacteria and is a lactose analog. Once it enters the cell, it will produce a long-term induction effect. Therefore, the induction efficiency of IPTG is high, and only a small amount (1mmol/L) is needed to achieve the ideal induction effect. Since IPTG is not metabolized, it specifically induces the expression of foreign proteins in the cell. Not affected by cell metabolism, the induction effect is stable. Lactose has a double effect. It can be used as a precursor of the inducer iso-lactose and a carbon source. During the induction process, it is very unstable. IPTG is suitable for experimental research due to its excellent stability. 3.IPTG can be effectively copied in the absence of lacY gene. 4. In the galactoside bond, sulfur is used instead of oxygen and the hydrolysis activity is lost. However, the affinity of thiogalactoside and homologous oxygen compounds are the same as the enzyme site. Although IPTG is not recognized by β-galactosidase, it is a very effective inducer of lac gene cluster. usage: IPTG is first prepared into a 23.83 mg/ml(100 mM) aqueous solution, filtered and sterilized and stored. Then, 100 μl of the above solution, 200 μl of X-Gal(20 mg/ml of dimethylformamide (DMF) solution) and 100 μl of Amp(100 mg/ml) were added to 100 ml of agar medium to prepare IPTG, X-Gal and Amp plate medium. When dna fragments are inserted into pUC series vectors (or other vectors with lacZ and Amp genes) and then transformed into lacZ deletion cells, the above IPTG, X-Gal and Amp plate culture media can be coated to conveniently select gene recombinants (white is gene recombinants with DNA insertion fragments) according to the blue-white color of the bacteria.
preparation of experimental materials	1. induced expression materials: (1) LB (Luria-Bertani)) medium: yeast extract (Yeast extract) 5g peptone (Peptone) 10g NaCl 10g agar (Agar) 1-2% distilled water (Distilled water) 1000ml pH 7.0 scope of application: escherichia coli (2) IPTG stock solution: 2g IPTG dissolved in 10 mL distilled water, 0.22 μm filter membrane filtration sterilization, sub-package into 1 mL/part,-20 ℃. (3) l× gel electrophoresis sample addition buffer: 50 mmol / L Tris-CI ( pH 6.8) 50 mmol / L DTT 2% SDS (electrophoresis grade) 0.1% bromophenol blue 10% glycerol 2, Escherichia coli inclusion body separation and protein purification materials: 1) enzymatic method (1) lysis buffer: 50 mmol / L Tris-CI ( pH 8.0) 1 mmol / L EDTA 100 mmol / LNaCI (2)50 mmol / L benzylsulfonyl fluoride (PMSF ). (3)10 mg/mL lysozyme. (4) deoxycholic acid. (5)1 mg/mL DNase I. 2) ultrasonic fragmentation (1) TE buffer. (2) 2 × SDS -PAGE gel electrophoresis sample addition buffer: 100 mmol / L Tris-HCI ( pH 8.0) 100 mmol / L DTT 4% SDS 0.2% bromophenol blue 20% glycerol
experimental scheme	1, induced expression of foreign genes: (1) digest vector DNA and target gene with appropriate restriction endonuclease. ( 2) connect the target gene and vector according to the connection step, and transform to the corresponding host bacteria. ( 3) screen out the transformed colonies containing recombinants, extract plasmid DNA for restriction endonuclease map, DNA sequence determination, after confirmation, proceed to the next step. ( 4) if the prokaryotic promoter of the expression vector is PL promoter, culture at 30 -32 ℃ for several hours to make OD600 of the culture solution reach 0.4-0.6, quickly raise the temperature to 42 ℃ and continue to culture for 3-5h; If the prokaryotic promoter of the expression vector is tac, etc., the bacteria are cultured at 37 ℃ for several hours to reach the logarithmic growth period, IPTG is added until the final concentration is 1 mmol / L. the culture is continued for 3-5h. ( 5) 1 mL of the above culture solution is taken, 1000g is centrifuged, 1 min is precipitated, after adding 100 μL polyacrylamide gel electrophoresis sample loading buffer, SDS -PAGE was used for detection. 2. separation of inclusion bodies of Escherichia coli and protein purification: lysis of bacteria common methods include: ① high temperature bead grinding method; ② high pressure homogenate; ③ ultrasonic crushing method; ④ enzyme dissolution method; ⑤ chemical infiltration, etc. The first three methods belong to mechanical crushing method, and methods ① and ② have been applied in industrial production, while the latter three methods are widely used in laboratory research. The following is an introduction to the experimental steps of the enzymatic method and the ultrasonic fragmentation method. Enzymatic method. Commonly used lysozyme; β-1, 3-glucanase; β-1, 6-glucanase; protease; chitanase; glycosase, etc. Lysozyme mainly acts on bacteria, while other enzymes have significant effects on yeast. The main steps are: 4 ℃, centrifugation at 5000rpm, 15 min, collecting the induced expression of bacterial culture solution (100 mL ). Abandon supernatant, add 3 mL lysis buffer per gram of wet bacteria, and suspend precipitation.
Uses	Commonly used molecular biology reagents, commonly used for blue and white spot screening and IPTG-induced protein expression in bacteria, etc. It is β-galactosidase and β-galactoporase Inducer, not hydrolyzed by β-galactoside, it is the substrate of thiogalactosyltransferase IPTG is often used in cloning experiments that require the induction of β-galactosidase activity. It is often used in combination with X-Gal or Bluo-Gal for blue-white screening of recombinant bacterial colonies, which can induce the expression of lac operon in Escherichia coli. IPTG binds to lacI repressor protein and changes its conformation to play a role in preventing the inhibition of the β-galactosidase encoding gene lacZ.

Last Update：2024-04-10 22:29:15

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