イソプロピル 1-チオ-β-D-ガラクトピラノシド 化学特性,用途語,生産方法
外観
白色, 結晶性粉末〜粉末又は塊
溶解性
水に易溶。水に溶ける。
用途
イソプロピル-β-チオガラクトピラノシド（Isopropyl β-D-1-thiogalactopyranoside）は、分子生物学で使用される試薬の一つである。通常は略してIPTGと呼ばれる。
この化合物はアロラクトースの類似体として用いられ、ラクトースオペロンの転写を誘導する。アロラクトースとは異なり、生体内では硫黄原子の部分で分解されないため、ラクトースオペロンは常に誘導された状態となる。
IPTGはラクトースリプレッサーに結合してその働きを阻害し、ラクトースを分解するβ-ガラクトシダーゼの発現を誘導する。クローニングにおいては、lacZ遺伝子の部分に目的の遺伝子が導入され、IPTGはその遺伝子の発現を誘導することになる。
用途
β-ガラクトシダーゼの誘導物質(inducer)。
説明
IPTG (also known as Isopropyl-β-D-Thiogalactopyranoside) is a molecular biology reagent that functions as an inducer of galactosidase activity by binding to and inhibiting the repressor. It is a molecular mimic of allolactose, a lactose metabolite that triggers transcription of the lac operon, and it is therefore used to induce protein expression where the gene is under the control of the lac operator. IPTG cannot be hydrolyzed or broken down by the E. coli therefore the concentration remains constant during cell replication. IPTG is often used at a final concentration ranging from 0.5-1.0 mM. X-Gal is often used with IPTG since it provides an easy visual method to monitor whether protein expression has occurred. X-Gal as the name implies contains a galactose group that once metabolized causes a bright blue color change.
化学的特性
White crystalline powder
使用
Isopropyl-β-D-thiogalactoside is used as a reagent in molecular biology. It is used as an effective β-galactosidase inducer of protein expression where the gene is under the control of the lac operator. It is used with 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside to identify the non-recombinant plasmid in cloning experiments by blue-white screen methodology.
製造方法
a synthetic method of isopropyl-β-D-thiogalactoside. The steps are as follows:
Step 1: Dissolve β-D-galactose pentaacetate in any organic solvent of methylene chloride, chloroform or 1,2-dichloroethane, and add Lewis acid (boron trifluoride ether, (Aluminum trichloride, zinc chloride) catalyst and potassium ethylxanthate or sodium ethylxanthate are reacted to obtain tetraacetylgalactose ethylxanthate after treatment.
Step 2: Dissolve tetraacetyl galactose ethyl xanthate, 2-bromopropane and sodium carbonate in methanol or ethanol at a temperature of 20°C to 80°C and react for 4-6 hours to obtain isopropyl group after treatment Isopropyl-β-D-thiogalactoside.
定義
ChEBI: Isopropyl beta-D-thiogalactopyranoside is an S-glycosyl compound consisting of beta-D-1-thiogalactose having an isopropyl group attached to the anomeric sulfur.
主な応用
IPTG (Isopropyl-β-D-thiogalactopyranoside) is a galactose analogue not recognized by β-galactosidase. IPTG is a non-metabolizable galactose analog that induces expression of the lac operon in Escherichia coli. It is a commonly used reagent in cloning procedures that require induction of b-galactosidase activity and is used in conjunction with X-Gal. It is commonly used in cloning procedures that require induction of β-galactosidase activity. It is also used in conjunction with X-Gal or Bluo-Gal in blue-white selection of recombinant bacterial colonies that induce expression of the lac operon in Escherichia coli. IPTG functions by binding to the lacI repressor and altering its conformation, which prevents the repression of the β-galactosidase coding gene lacZ.
一般的な説明
Inducer for β-galactosidase, an enzyme that promotes lactose utilization. Use in conjunction with X-Gal. Molecular Biology grade.
Stock solutions:
X-Gal: 20 mg/ml in DMF
IPTG: 20 mg/ml in water
Use 4:1 (X-Gal:IPTG)
Assay: ≥99%
Dioxane: None detected
Melting point: 110-114°C
Specific rotation [a]25/D (1%, water): -34.0 to -29.0°
Intended for laboratory and manufacturing use only. Not for drug, food, or household use.
イソプロピル 1-チオ-β-D-ガラクトピラノシド 上流と下流の製品情報
原材料
準備製品