^(15)N稳定同位素标记副溶血弧菌的培养基制备与培育  

Preparation of Culture Medium and cultivation of Vibrio Parahaemolyticus Labelled with ^(15)N Stable Isotope

作　　者：赵丽青[1] 王勇[1] 黄小华[1] 张晓良[1] 刘恩德[1] 胡炜[1] 孙宇 吴振兴[1] 贾俊涛[1] 李正义[1] 

机构地区：[1]山东出入境检验检疫局检验检疫技术中心,山东青岛266002 [2]贵州出入境检验检疫局,贵州贵阳550000

出　　处：《中国动物检疫》2018年第3期72-75,80,共5页China Animal Health Inspection

基　　金：国家质检总局科技计划项目(2013IK175;2015IK203;2016IK198;2014IK024)

摘　　要：为进行蛋白质组学的定量研究,研制一种^(15)N稳定同位素标记的副溶血弧菌培养基并进行^(15)N标记副溶血弧菌培育。方法包括连续多次转接培养副溶弧菌和利用LC-MS/MS验证标记的副溶血弧菌细胞中蛋白质氮原子的标记效率。所述培养基的配方为NH_4Cl(2 g/L)、葡萄糖(10 g/L)、KH_2PO_4(0.6 g/L)、KH_2PO_4(0.9g/L)、MgSO_4(0.2 g/L)、Na Cl(20 g/L)和蒸馏水(1L,pH 7.0~7.5);氮源的氮原子采用^(15)N进行标记,标记效率为91.5%。结果表明,该工艺简单合理,培养基组分简单、成本低,为工业化制备^(15)N标记副溶血弧菌培养基提供了新的制备方法。In order to conduct quantitative study of proteomics and to develop Vibrio parahaemolyticus culture medium labelled with 15N stable isotope,by the method of 15N marking Vibrio parahaemolyticus,including repeatedly transferring and cultivating Vibrio parahaemolyticus and verifng the labeling ef？ciency of nitrogen-atoms in the Vibrio parahaemolyticus bacterial cells by LC-MS/MS,the culture medium was obtained. The formula of the culture medium was：NH4Cl（2 g/L）,glucose（10 g/L）,KH2PO4（0.6 g/L）,KH2PO4（0.9 g/L）,MgSO4（0.2 g/L）,NaCl（20 g/L）and distilled water（1 L,pH 7.0-7.5）. The nitrogenous sourced nitrogen-atoms were labeled by 15N,and the the labeling ef？ciency was 91.5%. The results showed that the technology was simple and reasonable,and its components were simple and low cost. As a conclusion,it would provide a new method for preparing Vibrio parahaemolyticus culture medium labeled with 15N in industry.

关 键 词：15N 稳定同位素 副溶血弧菌 液相色谱-串联质谱 

分 类 号：S852.61[农业科学—基础兽医学]