摘要
为了探索DGAT1和DGAT2基因对宗地花猪脂肪沉积效率的影响,以宗地花猪为试验动物,针对基因DGAT1和DGAT2 mRNA序列设计6对shRNA序列,构建RNA干扰表达载体,将DGAT1和DGAT2的shRNA干扰载体分别转染至肝细胞中,采用荧光定量PCR筛选出最佳shRNA干扰载体,并测定最佳干扰组细胞代谢液中甘油三酯和总胆固醇含量变化。结果显示,所构建的6个shRNA干扰载体均能抑制肝细胞中目的基因的表达,肝细胞中DGAT1和DGAT2基因的最高抑制效率分别为78.45%和87.52%。转染干扰效率最佳的载体p GPU6/GFP/Neo-DGAT1-1和p GPU6/GFP/Neo-DGAT2-1后,肝细胞培养液中甘油三酯和总胆固醇含量均降低,两者差异不显著(P>0.05)。转染p GPU6/GFP/Neo-DGAT2-1的肝细胞培养液中甘油三酯含量显著低于空白组(P<0.05),DGAT1干扰载体的干扰效率略低于DGAT2干扰载体(P>0.05)。综上,抑制DGAT1和DGAT2基因的表达,可降低肝细胞培养液中甘油三酯和总胆固醇含量。
In order to explore the effects of DGAT 1 and DGAT 2 genes expression on fat deposition efficiency in pigs,six pairs of short hairpin RNA(shRNA)sequences were designed on the basis of mRNA of DGAT 1 and DGAT 2,and the Zongdihua pigs were used as experimental animals.After transfected into hepatocytes respectively,the best shRNA interference vectors for DGAT 1 and DGAT 2 mRNA in hepatocytes were selected by RT-qPCR,and the triglyceride and total cholesterol contents were tested in cell metabolic fluid.The results showed that all the shRNA interference vectors could inhibit the expression of target genes in hepatocytes,and then the highest inhibition efficiencies for DGAT 1 and DGAT 2 genes expression in hepatocytes were tested to be 78.45%and 87.52%,respectively with pGPU6/GFP/Neo-DGAT 1-1 and pGPU6/GFP/Neo-DGAT 2-1 vectors,however the contents of triglyceride and total cholesterol were decreased in cell metabolic fluid,but no significantly difference between them(P>0.05).The contents of triglyceride of hepatocyte metabolic fluid in pGPU6/GFP/Neo-DGAT 2-1 group was significantly lower than that of blank group(P<0.05).The interference efficiency of DGAT 1 was lower than that of DGAT 2(P>0.05).Inhibiting the expression of DGAT 1 and DGAT 2 could reduce the content of triglyceride and total cholesterol in the hepatocyte metabolic fluid.
作者
吴光松
李平
顾丽菊
林鹏飞
任丽群
杨蓉
黄维江
张芸
毛同辉
魏小红
田松军
燕志宏
WU Guangsong;LI Ping;GU Liju;LIN Pengfei;REN Liqun;YANG Rong;HUANG Weijiang;ZHANG Yun;MAO Tonghui;WEI Xiaohong;TIAN Songjun;YAN Zhihong(College of animal science,Guizhou University,Guiyang 550025,China;Mountain Plateau Animal Genetic Breeding and Reproduction of Ministry of Education Key Laboratory,Guizhou University,Guiyang 550025,China;Guizhou Provincial Animal Germplasm Measuring Center,Guiyang 550018,China;Guizhou Excellent Agricultural Valley Eco Industrial Co.Ltd,Guiyang 550001,China;Guizhou Agricultural Regional Economic Development Center, Guiyang 550001,China;Animal Husbandry Technology Promotion Station in Tongren City,Tongren 554300,China;Animal Husbandry Service Center in Ziyun,Anshun 550800,China)
出处
《河南农业科学》
CSCD
北大核心
2018年第8期123-127,共5页
Journal of Henan Agricultural Sciences
基金
贵州省科技厅农业公关项目(700712162104
700712121102)
