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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (15): 152-157.doi: 10.11924/j.issn.1000-6850.2011-3737

所属专题： 生物技术

• 生物技术科学 • 上一篇    下一篇

用SOE PCR方法获得新城疫病毒La Sota株全基因组cDNA

葛菁萍 宋姗姗 唐晓艳 高冬妮 张贺楠 楼庄伟 平文祥   

  • 收稿日期:2011-12-09 修回日期:2012-03-07 出版日期:2012-05-25 发布日期:2012-05-25
  • 基金资助:

    纤毛虫表膜内表面与射出体、皮层颗粒、色素颗粒的形态学研究;黑龙江省教育厅重点项目;黑龙江省科技攻关重大项目;科技创新人才研究专项资金项目优秀学科带头人;黑龙江大学高层次人才（创新团队）支持计划

Genome-wide cDNA of Newcastle Disease Virus Isolates Obtained from La Sota According to the SOE PCR Method

ping jingge   

  • Received:2011-12-09 Revised:2012-03-07 Online:2012-05-25 Published:2012-05-25

PDF (PC)

802

摘要：

为了获得新城疫病毒La Sota株全基因组cDNA。根据SOE PCR方法要求设计引物，对NDV La Sota株进行分段扩增，然后应用SOE PCR方法连接各片段，全基因连接可得到约15000 bp的扩增片段，所得到的产物为NDV La Sota株全基因组cDNA。结果表明：采用SOE PCR方法合成的基因序列经测序及酶切鉴定，与GenBank上登陆的NDV La Sota株全基因组序列基本一致。本研究为构建新城疫La Sota株的感染性cDNA奠定了物质基础；并为进一步研究NDV的生物学特性、结构与功能的关系，探讨影响NDV毒力的因素、及研制新型疫苗载体提供了可靠依据。

关键词: 反演检验, 反演检验

Abstract:

In order to obtain Newcastle disease virus genome-wide cDNA of La Sota strain through a series of primers splicing. According to SOE PCR method, it was required to design primer. As for NDV La Sota strains, first sectional amplification was carried out, and then applied the SOE PCR method to connect the fragments. Genome-wide connections could obtained about 15000 bp amplified fragments. It’s turned out that the resulting product was genome-wide cDNA of the NDV La Sota. The results showed that the gene sequences which synthesised by SOE PCR method was sequenced and digest by restriction enzyme. It’s turned out that the gene sequences were consistent with the expected results. This study laid the foundation for the construction of Newcastle disease La Sota strain’s infectious cDNA. It also provided a reliable guarantee for the further study of the biological properties of NDV, the relations between its structures and functions, and to explore the factors that affect NDV virulence, as well as the development of new vaccines.