Guan Li-zeng, Sun Yu-ping, Cai Jin-shun, Wu Han-dong, Yu Long-zheng, Zhang Yong-liang, Xi Qian-yun
Agriculture College, Yanbian University, Gongyuan Road, Yanji, 133000, China,
Transgenic Res. 2015 Jun;24(3):489-96. doi: 10.1007/s11248-015-9863-y. Epub 2015 Jan 21.
The aflatoxin-detoxifizyme (ADTZ) gene derived from Armillariella tabescens was cloned into parotid gland-specific expression vector (pPSPBGPneo) to construct the parotid gland-specific vector expressing ADTZ (pPSPBGPneo-ADTZ). Transgenic mice were generated by microinjection and identified by using PCR and Southern blotting analysis. PCR and Southern blotting analysis showed that total six transgenic mice carried the ADTZ gene were generated. RT-PCR analysis indicated that the expression of ADTZ mRNA could be detected only in parotid glands of the transgenic mice. The ADTZ activity in the saliva was found to be 3.72 ± 1.64 U/mL. After feeding a diet containing aflatoxin B1 (AFB1) for 14 days, the effect of ADTZ on serum biochemical indexes and AFB1 residues in serum and liver of mice were evaluated. The results showed that total protein and globulin contents in the test treatment (transgenic mice) produced ADTZ were significantly higher than that of the positive control, while alanine aminotransferase and aspartate aminotransferase activity in serum of the test treatment (transgenic mice) were remarkably lower compared to that of the positive control (P < 0.05). Moreover, AFB1 residues in serum and liver of the test treatment (transgenic mice) were significantly lower compared with that of the positive control (P < 0.05). These results in the study confirmed that ADTZ produced in transgenic mice could reduce, even eliminate the negative effects of AFB1 on mice.
将来源于亮菌的黄曲霉毒素解毒酶(ADTZ)基因克隆到腮腺特异性表达载体(pPSPBGPneo)中,构建表达ADTZ的腮腺特异性载体(pPSPBGPneo-ADTZ)。通过显微注射产生转基因小鼠,并利用PCR和Southern印迹分析进行鉴定。PCR和Southern印迹分析表明,共产生了6只携带ADTZ基因的转基因小鼠。RT-PCR分析表明,仅在转基因小鼠的腮腺中可检测到ADTZ mRNA的表达。唾液中的ADTZ活性为3.72±1.64 U/mL。在给小鼠喂食含黄曲霉毒素B1(AFB1)的饲料14天后,评估ADTZ对小鼠血清生化指标以及血清和肝脏中AFB1残留量的影响。结果显示,产生ADTZ的试验组(转基因小鼠)中的总蛋白和球蛋白含量显著高于阳性对照组,而试验组(转基因小鼠)血清中的丙氨酸氨基转移酶和天冬氨酸氨基转移酶活性与阳性对照组相比显著降低(P<0.05)。此外,试验组(转基因小鼠)血清和肝脏中的AFB1残留量与阳性对照组相比显著降低(P<0.05)。该研究中的这些结果证实,转基因小鼠产生的ADTZ能够减轻甚至消除AFB1对小鼠的负面影响。