CAJ | 학술논문

采用斑迹抽提法提取柞蚕微孢子虫(Nosema pernyi)基因组DNA,基因组DNA的琼脂糖凝胶电泳图谱中有大小约15 kb的清晰、完整条带.选用已报道的微粒子属16S rRNA基因的保守序列设计P1/P2和N1/N2 2对引物,对柞蚕微孢子虫基因组DNA进行PCR扩增,结果2对引物分别扩增出1条大小不同的特异条带,其中用引物N1/N2扩增可检测出稀释至1.5 ng的DNA模板.应用同样的PCR引物、体系和反应条件,可有效扩增出受感染柞蚕幼虫、成虫中的微孢子虫基因组DNA条带.该项检测技术有望应用于柞蚕微粒子病的早期诊断.
채용반적추제법제취작잠미포자충(Nosema pernyi)기인조DNA,기인조DNA적경지당응효전영도보중유대소약15 kb적청석、완정조대.선용이보도적미입자속16S rRNA기인적보수서렬설계P1/P2화N1/N2 2대인물,대작잠미포자충기인조DNA진행PCR확증,결과2대인물분별확증출1조대소불동적특이조대,기중용인물N1/N2확증가검측출희석지1.5 ng적DNA모판.응용동양적PCR인물、체계화반응조건,가유효확증출수감염작잠유충、성충중적미포자충기인조DNA조대.해항검측기술유망응용우작잠미입자병적조기진단.
The genomic DNA of Nosema pemyi was extracted by means of"disease-spot"method.The genomic DNA band with a length of around 15 kb was clear and intact in the agarose gel electrophoretogram.PCR amplifications using two pairs of primers(P1/P2 and N1/N2)designed in accordance with conserved regions of the reported Nosema 16S rRNA genes to amplify the genomic DNA showed that both primer pairs yielded one specific DNA band of different size respectively frOm the genomic DNA of Nosema pernyi.among which amplification with primers N1/N2 could detect 0.47 na template DNA.It is found that PCR amplification with the same primers,rection system and rection condition can detect the genomic DNA of Nosema pemyi in tussah silkworm larvae and moths effectively,showing high potential in ear-ly diagnosis of tussah silkworm pebrine disease.