CRISPR/Cas9介导的MSTN/FGF5基因编辑陕北白绒山羊粪便菌群分析

Fecal Microbial Flora Analysis in MSTN/FGF5 Gene-modified Shaanbei White Cashmere Goat Mediated by CRISPR/Cas9

为了从绒山羊粪便微生物群落变化的角度对MSTN/FGF5基因编辑陕北白羊绒山的自身健康和环境安全性进行研究,利用6种不同培养基分别对MSTN/FGF5基因编辑和野生型陕北白绒山羊粪便中的微生物进行培养、计数,并做菌群结构的显著性检验分析。结果表明:FGF5/MSTN基因编辑和野生型陕北白绒山羊F0代和F1代公羊和母羊新鲜粪样经LB琼脂、伊红美蓝琼脂、TPY琼脂、胆硫乳琼脂、肠球菌琼脂和甘露醇盐琼脂培养后微生物计数分别为,F0代:公羊6.35±0.68,5.89±0.25;5.32±0.10,5.27±0.34;6.64±0.57,6.71±0.35;5.03±0.53,5.32±0.14;4.12±0.13,4.15±0.21;5.34±0.18,5.25±0.25;母羊6.62±0.45,6.64±0.31;4.80±0.51,4.75±0.45;7.20±0.30,7.15±0.55;4.30±0.52,4.42±0.62;5.83±0.51,5.73±0.86;6.24±0.28,6.24±0.21。F1代:公羊6.92±0.12,7.08±0.04;4.02±0.64,3.99±0.51;6.32±0.15,6.25±0.30;5.21±0.28,5.17±0.20;3.66±0.50,3.88±0.67;4.06±0.50,3.97±0.46;母羊7.12±0.24,6.96±0.19;4.15±0.19,4.25±0.24;6.45±0.30,6.56±0.45;4.97±0.42,4.87±0.38;4.04±0.96,3.96±0.57;4.10±0.28,3.85±0.37。F0和F1代MSTN/FGF5基因编辑陕北白绒山羊与野生型陕北白绒山羊粪便中菌群结构之间不存在显著性差异(P>0.05)。本研究为FGF5/MSTN基因编辑陕北白绒山羊的安全性评价研究提供了试验数据和研究基础。

To study the health and environment safety of MSTN/FGF5 gene-modified Shaanbei Cashmere goats from the aspect of the microbe population change in goat feces, six kinds of mediums were used to culture fecal bacteria from MSTN/FGF5 gene-modified and wild-type goats, respectively, and the number of microbe was counted. The significance test analysis was also conducted. Results showed that microbial flora of feces,which were cultured on different mediums including Luria-Bertani, Eosin methylene blue agar and TPY plates,Bile Esculin Azide, DHL agar, Mannitol salt selective, were as follows： F0, male, 6.35±0.68, 5.89±0.25; 5.32±0.10, 5.27±0.34; 6.64±0.57, 6.71±0.35; 5.03±0.53, 5.32±0.14; 4.12±0.13, 4.15±0.21; 5.34±0.18, 5.25±0.25;female 6.62±0.45, 6.64±0.31; 4.80±0.51, 4.75±0.45; 7.20±0.30, 7.15±0.55; 4.30±0.52, 4.42±0.62; 5.83±0.51,5.73±0.86; 6.24±0.28, 6.24±0.21. F1： male 6.92±0.12, 7.08±0.04; 4.02±0.64, 3.99±0.51; 6.32±0.15, 6.25±0.30; 5.21±0.28, 5.17±0.20; 3.66±0.50, 3.88±0.67; 4.06±0.50, 3.97±0.46; female 7.12±0.24, 6.96±0.19; 4.15±0.19, 4.25±0.24; 6.45±0.30, 6.56±0.45; 4.97±0.42, 4.87±0.38; 4.04±0.96, 3.96±0.57; 4.10±0.28, 3.85±0.37.Results indicated that in comparison with the control goats, microbe population of various species in feces of MSTN/FGF5 gene-modified goats remained unchanged（P〈0.05）. In summary, this study provides the experimental data and a research basis for the safety evaluation of MSTN/FGF5 gene-modified Shaanbei Cashmere goat.