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家蚕微孢子虫原位杂交诊断技术研究
Studies on Detection of Nosema bombycis by Hybridization in situ
【摘要】 根据家蚕微孢子虫rRNA基因高度保守的特点,设计合成了1对PCR引物,从家蚕微孢子虫基因组DNA上扩增出1个12kb片段,用同位素标记后作为检测家蚕微孢子虫的特异性探针。采用原位杂交的方法,对家蚕卵及感染家蚕微孢子虫4、6、10d的幼虫进行原位杂交检测。实验结果表明,该方法可以从家蚕单粒卵内检测到296粒孢子,对感染家蚕微孢子虫4、6、10d的幼虫均有阳性杂交信号出现。
【Abstract】 For the purpose of detecting Nosema bombycis by hybridization in situ, the probe(1.2 kb) was obtained by PCR from rRNA gene of Nosema bombycis and labeled by random primer synthesis technique with ~32 P-dATP.The result showed that at least 296 spores were detected from single silkworm egg,and the spores were also detected from the larvae infected by Nosema bombycis for 4 days ,6 days and 10 days respectively.
【关键词】 家蚕微孢子虫;
家蚕卵;
家蚕幼虫;
原位杂交;
诊断;
【Key words】 Nosema bombycis; Bombyx mori eggs; Bombyx mori larva; Hybridization in situ; Diagnosis;
【Key words】 Nosema bombycis; Bombyx mori eggs; Bombyx mori larva; Hybridization in situ; Diagnosis;
【基金】 国家社会公益研究专项[国科发财字(2002)7号];国家茧丝绸发展风险基金[国茧协办(2002)30号]。
- 【文献出处】 蚕业科学 ,Acta Sericologica Sinica , 编辑部邮箱 ,2005年01期
- 【分类号】S884.21
- 【被引频次】23
- 【下载频次】189