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发明授权
CN101864445B 携带分子标记的猪瘟病毒C株感染性cDNA载体的构建方法 有权 许可

携带分子标记的猪瘟病毒C株感染性cDNA载体的构建方法
摘要：
本发明涉及病毒抗体研究，旨在提供一种携带分子标记的猪瘟病毒感染性cDNA载体的构建方法。包括：(1)用生理盐水稀释猪瘟活疫苗至1头份/ml作为实验材料，抽提总RNA后，根据设计的6对引物，RT-PCR分段扩增相应的6个cDNA片段；(2)cDNA片段F1和F5中相应分子标记的引入；(3)全长cDNA克隆所需质粒的改造；(4)全长cDNA载体的构建。通过本发明可获得携带分子标记的猪瘟病毒兔化弱毒疫苗C株的感染性cDNA载体pA-FL22。由该载体获得的RNA电转猪瘟病毒的易感细胞，拯救具有感染性的子代病毒，且该子代病毒能稳定遗传上述构建中引入的分子标记。利用该感染性cDNA载体及子代病毒的拯救技术体系，可为深入研究该病毒的复制机制、致病机理以及新型标记疫苗开发奠定基础。
摘要（英）：

The invention relates to a virus antibody research and provides a method for constructing a hog-cholera virus infectious cDNA carrier having a molecule mark. The method comprises the following steps of: (1) diluting a hog-cholera live vaccine to 1 portion per milliliter as the experimental material, after extracting the head RNA, amplifying 6 corresponding cDNA fragments through RT-PCR section according to designed 6 pairs of primers, (2) leading in the corresponding molecule marks from F1 and F5 of cDNA fragments, (3) reforming plasmids required by overall length cDNA cloning, and (4) constructing the overall length cDNA carrier. A hog-cholera virus lapinized vaccine C stem infectious cDNA carrier pA-FL22 having a molecule mark can be acquired through the method of the invention. The permissive cells of RNA electro transferred hog-cholera virus acquired by the carrier can save the infectious progeny virus which can stably inherit the molecule mark led in the construction. By using the saving technique system of the infectious cDNA carrier and the progeny virus, the copy stem and the pathopoiesia reason of the virus can be deeply researched and the foundation for developing new marked vaccines can be settled.

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