黄曲霉毒素解毒酶在小鼠肠道中的特异性表达  

Intestine-specific expression of the aflatoxin-detoxifizyme in mice

作　　者：关立增[1,2] 娄安钢 刘月[1] 胡楠 蔡锦顺[1] 

机构地区：[1]延边大学农学院,吉林延吉133002 [2]临沂大学农林科学学院,山东临沂276005

出　　处：《中国兽医学报》2018年第2期378-382,共5页Chinese Journal of Veterinary Science

基　　金：吉林省教育厅资助项目

摘　　要：通过筛选出黄曲霉素解毒酶（ADTZ）基因片段，采用鼠的黏蛋白启动子（MUC）成功构建了ADTz基因肠道特异性表达载体（MUC—ADTZ-LV），并利用原核注射法制备了肠道特异性表达ADTZ转基因小鼠。PCR和Southernblot鉴定结果显示，成功制备了6只携带肠道特异性表达ADTZ基因转基因小鼠。RT_PcR结果显示，ADTZ基因仅在小鼠肠道中进行了特异性表达。肠液中ADTz的活性为（4．62±1．54）U／mL。在饲料中添加黄曲霉毒素并对小鼠进行14d的喂养试验，之后对血清生化指标及小鼠血清和肝脏中黄曲霉毒素的残留进行了测定。结果表明，转基因组血清中总蛋白（TP）和球蛋白（GLP）的含量显著高于阳性对照组，而转基因组血清中的丙氨酸氨基转移酶（ALT）和天冬氨酸氨基转移酶（AST）的含量显著低于阳性对照组（P〈0．05）。转基因组小鼠血清和肝脏中黄曲霉毒素的残留量显著低于阳性对照组小鼠血清和肝脏中黄曲霉毒素的残留量。本试验在转基因小鼠的肠道中成功生产了ADTZ，并降低了饲料中黄曲霉毒素对小鼠毒性的影响。In this study,the ADTZ gene fragment was selected and the ADTZ intestine tissue-spe- cific expressing vector （MUC-ADTZ-LV） was successfully constructed by using the mouse mucin protein （MUC） promoter, and the transgenic mice expressing ADTZ in the intestine tissue was successful prepared by microinjeetion method. PCR and Southern blot analysis showed that 6 transgenic cloned mice expressing specifically ADTZ in the intestinal tissue were obtained. RT- PCR analysis indicated that the expression of ADTZ could be detected only in the intestine tissue of the transgenic mice. The ADTZ activity in the intestinal juice was found to be （4.62 ± 1.54 ） U/mL. After feeding a diet containing aflatoxin （AFT） for 14 d,the effect of ADTZ on serum bio- chemical indexes and AFT residues in serum and liver of mice were detected. The results showed that total protein （TP） and globulin （GLB） contents in the transgenic mice produced ADTZ were significantly higher than that of the positive control,while alanine aminotransferase （ALT） and aspartate aminotransferase （AST） activity in serum of the transgenie mice were remarkably lower compared to that of the positive control （P〈0.05）. Moreover,AFT residues in serum and liver of the transgenic mice were significantly lower compared to that of the positive control （P〈0.05）. These results in the study confirmed that ADTZ produced in the intestine tissue of the transgenic mice could reduce the negative effects of AFT on mice.

关 键 词：肠道组织 黄曲霉毒素解毒酶 转基因小鼠 

分 类 号：S859.87[农业科学—临床兽医学]