番鸭VIPR1和miR-317以及MAP3K7和miR-244靶向关系的研究  

Verification of Targeting Relationship Between VIPR1 and miR-317 and MAP3K7 and miR-244 in Muscovy Duck

作　　者：李丽[1] 辛清武[1] 章琳俐[1] 缪中纬[1] 朱志明[1] 梁阿政 郑嫩珠[1] LI Li;XIN Qingwu;ZHANG Linli;MIAO Zhongwei;ZHU Zhiming;LIANG Azheng;ZHENG Nenzhu(Fujian Key Laboratory of Animal Genetics and Breeding,Institute of Animal Husbandry and Veterinary Medicine,Fujian Academy of Agricultural Sciences,Fuzhou,Fujian 350013;Zhangzhou Changlong Agriculture and Animal Husbandry Co.,Ltd.,Zhangzhou,Fujian 363100)

机构地区：[1]福建省农业科学院畜牧兽医研究所,福建省畜禽遗传育种重点实验室,福建福州350013 [2]漳州昌龙农牧有限公司,福建漳州363100

出　　处：《中国家禽》2024年第6期15-19,共5页China Poultry

基　　金：福建省省属公益类项目(2020R10260016);福建省自然科学基金项目(2021J01483);福建省农业科学院英才项目(YC20210046)。

摘　　要：试验旨在验证番鸭就巢相关miRNA与靶基因靶向关系。试验利用miRanda、PITA和RNAhybrid三个软件的交集预测miR-317和miR-244靶基因,构建VIPR1-pmirGLO/MAP3K7-pmirGLO野生型和突变型双荧光素酶重组载体,将miR-317/VIPR1-Wt和miR-244/MAP3K7-Wt分别共转染至鸭胚成纤维细胞,检测双荧光素酶活性。结果显示:与miR-NC组相比,miR-317/VIPR1-Wt共转染组相对荧光值显著降低(P<0.05);VIPR1突变后,miR-NC组与miR-317组的相对荧光值不显著(P>0.05),说明突变后完全抑制miR-317对VIPR1的结合作用,VIPR1与miR-317之间存在相互结合作用。与miR-NC组相比,miR-244/MAP3K7-Wt共转染组相对荧光值无显著性差异(P>0.05);MAP3K7突变后,miR-NC组与miR-244组的相对荧光值差异不显著(P>0.05),表明MAP3K7与miR-244之间不存在相互结合作用。研究提示,miR-317与VIPR1存在确切靶向结合位点,即VIPR1是miRNA miR-317的靶基因,而miR-244与MAP3K7不存在靶向关系。The aim was to verify the relationship between broodiness-related miRNA and target gene targeting in Muscovy ducks.The intersection of miRanda,PITA and RNAhybrid software were used to predict the target genes of miR-317 and miR-244.Double luciferase recombinant vectors of VIPR1-pmirGLO/MAP3K7-pmirGLO wild-type and mutant were constructed,and miR-317/VIPR1-Wt and miR-244/MAP3K7-Wt were co-transfected into duck embryo fibroblasts to detect the double lucifer⁃ase activity.The results showed that the relative fluorescence value of miR-317/VIPR1-WT co-transfection group decreased sig⁃nificantly(P<0.05)compared with the miR-NC group,while there was no significant change in the relative fluorescence value of miR-NC group and miR-317 group after VIPR1 mutation(P>0.05),which suggested that the binding effect of miR-317 on VIPR1 was completely inhibited after mutation,indicating an interaction between miR-317 and VIPR1.The results showed that there was no significant difference in the relative fluorescence values of miR-244/co-transfection group compared with the miRNC group(P>0.05).Moreover,the relative fluorescence values of miR-NC group and miR-244 group were not significant after MAP3K7 mutation(P>0.05),indicating that there was no interaction between miR-244 and MAP3K7.According to the findings,exact targeting binding site was identified between VIPR1 and miR-317,indicating that VIPR1 was the target gene of miR-317.However,MAP3K7 and miR-244 did not have a targeting relationship.

关 键 词：番鸭 就巢 miR-317 miR-244 MAP3K7 VIPR1 

分 类 号：S834.2[农业科学—畜牧学]