关键词: 绵羊, GDF9基因, G1突变, 可视化检测, ARMS, SYBR Green Ⅰ

Abstract: In order to establish a rapid and visual method for detecting the G1 mutation of growth differentiation factor 9 gene ( GDF9 ) in sheep, an amplification refractory mutation system(ARMS) specific primer pair for the G1 mutation of the GDF9 gene was designed,and additional mismatches at the 2nd to 4th bases at the 3′ end of the downstream primers were designed respectively to screen for primer pair with the best specificity.The G1 mutation of the GDF9 gene was visually detected by combining the improved ARMS with the nucleic acid dye SYBR Green Ⅰ.The results showed that the improved ARMS specific primers could distinguish between mutant genotype and wild type.The G1 mutation was visually detected by integrating the improved ARMS and SYBR Green Ⅰ.It was found that the known mutant genotype sample showed bright green,while the known wild type sample showed orange yellow,and the color of the two changed significantly.Twenty five small tail Han sheep samples were detected by the established method for visually detecting the G1 mutation of the GDF9 gene in sheep,and the results revealed that the method could distinguish the wild type and mutant genotype of the G1 mutation of the GDF9 gene in sheep accurately.The detection results were consistent with the sequencing results,and the accuracy was as high as 100%.Therefore,the established visual detection method can be used to detect the G1 mutation of the GDF9 gene in sheep.

Key words: Sheep, GDF9 gene, G1 mutation, Visual detection, ARMS, SYBR Green Ⅰ