| 摘要: |
| 于1997年5月在广东省珠海市采集斑马鱼和豹纹斑马鱼,采用热休克方法研究阻止第二极体排放诱导异源三倍体斑马鱼的适宜条件。按正交实验方案组合诱导参数,结果表明:在卵受精后2min,采用39℃持续处理2min,三倍化率可达53.8%,原肠期相对存活率为91.0%,尾芽期相对存活率为91%,孵化期相对存活率为91.1%。本研究分析了热休克处理的参数与三倍体出现率和胚胎相对存活率的关系,并得出诱导参数中起始休克时间为重要因素,其次为持续时间和休克温度。本研究为构建人工三倍体模型奠定了基础,并对三倍体的应用前景进行了讨论。 |
| 关键词: 基因组操作 异源三倍体 斑马鱼 正交实验 |
| DOI：10.11693/hyhz200005001001 |
| 分类号: |
| 基金项目:国家自然科学基金重点资助项目，39823003号 |
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| STUDIES ON THE INDUCTION OF ALLOTRIPLOID USING HEAT SHOCK IN ZEBRAFISH, BRACHYDANIO FRANKEI AND BRACHYDANIO RERIO |
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WU Yu-ping1, YE Yu-zhen2, WU Qing-jiang2
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1.Institute of Genetics, Maternal and Child Health Hospital of Zhuhai, Zhuhai519000;2.Institute of Hydrobiology, The Chinese Academy of Sciences, Wuhan430072
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| Abstract: |
| Brachydanio frankei and Brachydanio rerio were collected from the coastal waters of Zhuhai, Guagdong Province in May 1997. Temperature treatment method is utilized to study induction of allotriploid zebrafish. The present research elaborates an appropriate condition for production of allotriploid zebrafish by using heat shock with the retention of the second polar body. From the orthogonal composite design of L9 (33), an effective procedure for inducing triploid is the treatment of eggs at 39°C, 2 min after insemination for 2 min duration, which results in 53.8% triploidy embryos, 91.0%, 91.0% and 90.1% relative survival at gastrula stage, tail bud stage and hatching stage, respectively. The interrelationship between the percentage of triploid and survival with different inducing parameters are analyzed. It leads to conclusions that the most important factor for triploidy induction is initiative time of shock, the second is duration time and the third is shock temperature. |
| Key words: Genome manipulation, allotriploid, zebrafish, orthogonal test |
