| DPI染色显示黄姑鱼NOR的方法研究 |
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| 引用本文： | 陈紫瑩,;王浩华,;王凯武,;刘贤德,;王志勇,;蔡明夷.DPI染色显示黄姑鱼NOR的方法研究[J].厦门水产学院学报,2014(3):161-167. |
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| 作者姓名： | 陈紫瑩 ;王浩华 ;王凯武 ;刘贤德 ;王志勇 ;蔡明夷 |
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| 作者单位： | [1]农业部东海海水健康养殖重点实验室,福建厦门361021; [2]集美大学水产学院,福建厦门361021 |
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| 基金项目： | 国家自然科学基金项目（31272653）;2013年度福建省大学生创新性实验计划项目 |
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| 摘 要： | 研究了DPI染法显示黄姑鱼NOR的条件,并比较了DPI染法与银染法差别显示的位置、数目和直径.综合研究结果,DPI染法显示黄姑鱼NOR的程序可以归纳为：制片在60℃恒温箱中老化3～6h,接着在体积分数为80％的甲酰胺/2×SSC中65℃处理4min后,再用系列乙醇脱水,放在60℃下烘片5～10 min,然后滴加含有1μg/mL碘化丙啶（PI）的封片剂,盖上盖玻片,用指甲油封片.其中,热甲酰胺处理和PI染色是必须步骤;制片的老化和烘片虽非必须步骤,但可以提高差别显示的对比度.由DPI染法和银染法的对比试验结果显示,两种方法差别显示区域的位置一致,数目分布相当,直径没有显著性差异.可见,DPI染法可以替代银染法用于显示黄姑鱼间期核和中期染色体的NOR.由数目比较试验结果显示,DPI染法组仅个别间期核（0.81％）观察到3个NOR,而银染组观察到3个以上的间期核达3.51％.这表明,DPI染法比银染法有更高的特异性,更适合黄姑鱼群体NOR多态性研究.此外,DPI染法还可与FISH联用,同时定位NOR和其他基因位点.
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| 关 键 词： | 黄姑鱼 染色体 核仁组织区域 |
The Procedure of DPI Staining to Display Nucleolar Organizer Regions in Yellow Croaker （Nibea alibiflora） |
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| Institution: | CHEN Zi-ying, WANG Hao-hua, WANG Kai-wu, LIU Xian-de, WANG Zhi-yong, CAI Ming-yi ( 1. Key Laboratory of Helthy Maricuhure for the East China Sea, Ministry of Agriculture, Xiamen 361021, China; 2. Fisheries College, Jimei University, Xiamen 361021, China) |
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| Abstract: | The conditions for display nucleolar organizer regions （NORs） with propidium iodide （DPI） staining in yellow croaker （Nibea alibiflora） were investigated. The location, number and diameter of regions displayed by DPI staining and silver staining were compared. According to the results, the procedure of DPI staining for displaying NOR in yellow croaker could be summarized as follow ： age the slides at 60 ℃ for 3 - 6 h ; treat the slides in 65 ℃ and 80% volume fraction forrnamide （FA） / 2 × SSC for 4 min, followed with dehydration with series of ethanol ; bake the slides at 60 ℃ ; add anti - fade solution containing 1 μg/mL PI, cover it with a cover glass, and then seal with snail oil. Wherein, the treatment with FA and the PI staining were necessary steps; while the aging and the baking were unnecessary steps, but were able to improve the contrast between NOR and the background. Comparison between DPI staining and silver staining showed that the locations, numbers and diameters of NOR displayed with two methods had not significant difference.However, less ultra - number NORs were observed with DPI staining than with silver staining, suggesting higher specificity for DPI staining. Above all, DPI staining could replace silver staining to display NOR in yellow croaker, with several virtues, such as high specificity, easy handling. In addition, DPI could be used with FISH to map NOR and other loci simultaneously. |
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| Keywords: | Nibea alibiflora chromosome nucleolar organizer region |
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