Method for preparing 15N stable isotope labeled 5-methyldeoxycytidine

• Main Authors: ZENG YANYAN, ZHU DAN, FAN HONGBO, YUAN MEIYI, LIN BOJI, HU JUNJIE, LAO ZHILANG, LIANG ZHIHUI, LI YANLIANG, WU KANGMING
• Format: Patent
• Language: Chinese; English
• Published: 10.07.2018
• Subjects: APPARATUS THEREFOR; BEER; BIOCHEMISTRY; CHEMISTRY; DERIVATIVES THEREOF; ENZYMOLOGY; FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE; GENERAL METHODS OF ORGANIC CHEMISTRY; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; NUCLEIC ACIDS; NUCLEOSIDES; NUCLEOTIDES; ORGANIC CHEMISTRY; PROCESSES USING MICROORGANISMS; SPIRITS; SUGARS; VINEGAR; WINE

The invention relates to a method for preparing 15N stable isotope labeled 5-methyl deoxycytidine. The method comprises the following steps of step 1, selecting a strain; step 2, preparing a glucose culture medium containing 15N ammonium chloride and a liquid activation culture medium; step 3, culturing the strain selected in step 1 in two culture media prepared in step 2; step 4, extracting and preserving bacterial genomic DNA; step 5, conducting a CviPI DNA methylase catalytic reaction; step 6, conducing genomic DNA hydrolysis, and conducting separation and purification of 15N 5-methyl deoxycytidine. The method utilizes microbes to synthetize the 15N stable isotope labeled 5-methyl deoxycytidine, 15N is easy to label and has high abundance, process is simple, and preparation is convenient. 本发明涉及种N稳定性同位素标记5-甲基脱氧胞苷的制备方法，包括以下步骤：步骤1、选用菌株；步骤2、制备含有N氯化铵的葡萄糖培养基和液体活化培养基；步骤3、将步骤1所选用的菌株在步骤2制备两种培养基中培养；步骤4、细菌基因组DNA与细菌基因组DNA提取和保存；步骤5、CviPI DNA甲基化酶催化反应；步骤6、基因组DNA水解，N 5-甲基脱氧胞苷分离纯化。本发明利用微生物合成N稳定性同位素标记5-甲基脱氧胞苷，N容易标记且丰度高，工艺简单