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新城疫病毒F_(48)E_8株cDNA文库的构建 被引量:3

CONSTRUCTION OF cDNA LIBRARIES OF A CHINESE VIRULENT NEWCASTLE DISEASE VIRUS STRAIN F_(48)E_8
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摘要 以SPF鸡胚繁殖我国标准新城疫病毒F_(48)E_8强毒株,经蔗糖密度梯度超速离心纯化病毒。再以酚-SDS法提取病毒基因组RNA,作为反应模板、采用Promega公司商品试剂盒合成双股cDNA。以同聚物加尾的方法将cDNA克隆到质粒pGEM3Zf(一)中,经AIX平板筛选,限制性内切酶分析和Digoxigenin标记的核酸探针检测,共获得插入外源片段大小在0.6~4.8kb的阳性克隆75个。 The virulent strain F_(48)E_8 of Newcastle disease virus(NDV)was grown in SPF chicken eggs and purified bycentrifugation in a sucrose gradient,then extracted with phenol-SDS to gain genomic RNA that was used as template togenerate cDNA with RiboClone cDNA synthesis system of Promega Corporation. The two-strand cDNA was tailed with oli-go(dC)and annealed to plasmid pGEM 3Zf(一)which was cut with PstI and tailed with oligo(dG ). The recombinantswere transformed into comptent E. Coli., and 75 positive clones which inserted 0.6~4. 8kb cDNA fragments werescreened with AIX plates and dot-blot hybridization by Digoxigenin labeled probes.
出处 《江苏农学院学报》 CSCD 1996年第4期67-69,共3页 Jiangsu Agricultural Research
基金 江苏省自然科学基金 江苏省教委自然科学基金
关键词 新城疫 病毒 文库 鸡病毒 NDV F_(48)E_8 strain cDNA library
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参考文献1

  • 1宋长绪,95全国禽病分子生物学研讨会论文集,1995年

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