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The Lifespan Test of Drosophila   

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摘要:主要介绍果蝇寿命测试的实验方法,以便于研究不同基因与果蝇寿命的关联。

关键词: 果蝇, 寿命, 测试

材料与试剂

  1. 棉球
  2. 试管
  3. 果蝇食物
  4. 60只雄果蝇和60只处女果蝇

仪器设备

  1. 培养箱

软件

  1. Prism 6

实验步骤

注:

1)
用果蝇作寿命检测时一个的原则就是要保证你所感兴趣的基因突变系品系的遗传背景与野生型对照一致。做寿命检测前,所用的基因突变果蝇需与野生型对照已回交6~8代以上。
2)
如果突变株基因确实跟寿命有关系,那么还应该做好rescue实验,即在突变株基础上构建转基因果蝇用于突变基因功能的补回,防止所观察到的寿命影响效应是由于突变体改造时基因组存在的不特异敲除所导致的;除了全身性的过表达还要用不同组织特异性启动子如Gal4系统检测该基因在哪些部位 (如大脑、脂肪体、血淋巴等) 可以把突变对寿命的效应消除,以确定目标基因在哪个组织行使功能。

  1. 25 °C,60%湿度条件培养的果蝇,挑60只雄果蝇和60只处女果蝇,养3~5天。
  2. 每管食物里放3只雄果蝇和3只处女果蝇,准备10~20管。
  3. 5~6天后倒掉亲本果蝇。
  4. 10天左右开始出成虫,每天固定时间点挑一轮雄果蝇,每管放20只,标明名字、日期和只数,挑够200只即可。
  5. 每间隔一天换一次食物,并记录果蝇存活情况。
    注:
    1)
    换食物的时候一定要仔细,尽量避免果蝇飞走或者被棉球压死。
    2)
    每次换食物前要将食物预热到25 °C,并且保证没有水汽和染菌。
  6. 将所记录情况输入软件Prism 6进行统计分析。

数据分析

每组果蝇寿命测试的实验需要独立重复三次,每次寿命测试的实验都有野生型果蝇寿命测试作为对照,三次独立重复实验的median lifespan应该保持一致,如果相差太大,对照组实验肯定不可以作为参考,需要考虑野生型对照组果蝇的状态,比如果蝇生长的温度,湿度,食物及环境洁净度等因素。

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Copyright: © 2019 The Authors; exclusive licensee Bio-protocol LLC.
引用格式:邓娟, 刘南. (2019). 果蝇寿命测试. Bio-101: e1010255. DOI: 10.21769/BioProtoc.1010255.
How to cite: Deng, J and Liu, N. (2019). The Lifespan Test of Drosophila. Bio-101: e1010255. DOI: 10.21769/BioProtoc.1010255.
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