褪黑素合成酶AANAT/ASMT基因过表达绵羊生物安全评价研究

doi:10.11843/j.issn.0366-6964.2020.07.009

摘要/Abstract

摘要： 旨在研究乳腺过表达褪黑素合成酶基因AANAT和ASMT（HIOMT）绵羊的生物安全性，基于本实验室前期建立的乳腺过表达褪黑素合成酶基因AANAT和ASMT（HIOMT）绵羊模型，追踪阳性转基因绵羊的生长性状数据，分析血液及尿液生理生化指标，对肠道微生物、乳中褪黑素水平及乳成分进行检测。结果表明：1）阳性转基因绵羊0、6和12月龄的体重、体长、身高和胸围4项生长指标与普通绵羊均无显著差异（P>0.05）；2）阳性绵羊血液总蛋白、白蛋白、球蛋白、胆固醇含量和各类型细胞数量以及尿液中亚硝酸盐、蛋白质和葡萄糖含量等指标均属正常，且与普通绵羊均无显著差异（P>0.05）；3）菌群测序结果表明，阳性绵羊及普通绵羊肠道微生物群落组成及优势菌群均无显著差异（P>0.05）；4）转基因绵羊血液褪黑素表达水平与普通绵羊无显著差异（P>0.05），夜间褪黑素水平显著高于白天（P<0.05）；乳中褪黑素水平极显著高于普通绵羊（P<0.01），乳糖含量显著高于对照羊（P<0.05），体细胞数极显著低于普通绵羊（P<0.01）。综上所述，乳腺过表达AANAT和ASMT（HIOMT）转基因绵羊的生长发育、诸多生理生化指标、肠道微生物菌群等与对照组绵羊相比均无显著差异。此外，转基因绵羊乳中褪黑素和乳糖含量较高，体细胞数含量较低，可能是乳腺中褪黑素发挥抗炎作用，降低了乳中体细胞数。

关键词: 绵羊, 转基因, AANAT, ASMT, 安全评价

Abstract: The purpose of this experiment was to study the biological safety of sheep with melatonin synthase genes AANAT and ASMT (HIOMT) over-expressed in mammary gland. Based on the sheep models with AANAT and ASMT (HIOMT) over-expressed in mammary gland established in the previous study, we tracked the growth traits of positive transgenic sheep, analyzed the physiological and biochemical indexes of the blood and urine samples, and detected the intestinal microorganisms, melatonin levels in milk and milk composition. The results showed that:1) The body weight, body length, height and bust of positive transgenic sheep at 0, 6 and 12 months of age were not significantly different from those of ordinary sheep (P>0.05); 2) The total protein levels, albumin, globulin and cholesterol levels in blood of positive transgenic sheep as well as the number of various types of cells were not significantly different from those of ordinary sheep(P>0.05). Meantime, the levels of nitrite, protein, glucose and some other factors in urine were normal and not significantly different from those of ordinary sheep (P>0.05); 3) The sequencing results of microbiota showed that there was no significant difference in the composition of intestinal microbial communities and dominant bacteria between positive transgenic sheep and ordinary sheep(P>0.05); 4) The expression level of melatonin in blood of positive transgenic sheep was not significantly different from that of ordinary sheep (P>0.05), the level of melatonin at night was significantly higher than that at daytime (P<0.05); while the level of melatonin in milk of positive transgenic sheep was extremely significantly higher than that of ordinary sheep (P<0.01), the lactose content was significantly higher than that of ordinary sheep (P<0.05), in addition, the number of somatic cells of positive transgenic sheep was extremely significantly lower than that of ordinary sheep (P<0.01). In summary, the growth performance, many physiological and biochemical indexes and intestinal microbiota of positive transgenic sheep with AANAT and ASMT (HIOMT) over-expressed in the mammary gland were not significantly different compared with the ordinary sheep. Furthermore, the content of melatonin and lactose in milk of positive transgenic sheep were higher and the number of somatic cells was lower, which might be that melatonin in the mammary gland played an anti-inflammatory effect, reducing the number of somatic cells in the milk.

Key words: sheep, transgene, AANAT, ASMT, safety evaluation

中图分类号:

S826.2

杨爱玲, 李广栋, 吴昊, 姬鹏云, 张鲁, 张金龙, 张效生, 阿布力孜·吾斯曼, 连正兴, 刘国世. 褪黑素合成酶AANAT/ASMT基因过表达绵羊生物安全评价研究[J]. 畜牧兽医学报, 2020, 51(7): 1563-1572.

YANG Ailing, LI Guangdong, WU Hao, JI Pengyun, ZHANG Lu, ZHANG Jinlong, ZHANG Xiaosheng, WUSIMAN Abulizi, LIAN Zhengxing, LIU Guoshi. Study of Biosafety Evaluation on Melatonin Synthase AANAT/ASMT Overexpressed Sheep[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(7): 1563-1572.

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