| 家蚕浓核病毒中国(镇江)株基因组的克隆及重组质粒转染家蚕对浓核病毒的拯救 |
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| 引用本文： | 付艳红,唐顺明,覃光星,刘挺,郭锡杰.家蚕浓核病毒中国(镇江)株基因组的克隆及重组质粒转染家蚕对浓核病毒的拯救[J].蚕业科学,2008,34(3). |
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| 作者姓名： | 付艳红 唐顺明 覃光星 刘挺 郭锡杰 |
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| 作者单位： | 江苏科技大学生物与环境工程学院,江苏镇江,212018;中国农业科学院蚕业研究所,江苏镇江,212018;中国农业科学院蚕业研究所,江苏镇江,212018 |
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| 基金项目： | 国家重点基础研究发展计划(973计划) |
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| 摘 要： | 家蚕浓核病毒中国(镇江)株(BombyxmoriDensovirus Zhenjiang Strain,BmDNV-ZJ)包含VD1和VD2共2种基因组DNA。以BmDNV-ZJ感染家蚕中肠总DNA为模板,采用PCR方法扩增得到2种DNA,并将其克隆到质粒pGEM-T构建了重组质粒pGEM-VD1和pGEM-VD2。采用DEAE-dextran转染技术,将2种重组质粒分别导入家蚕幼虫体内,能使家蚕幼虫发病,免疫双向扩散和免疫酶组化法检测都能检出阳性反应,但转染pGEM-VD2重组质粒的家蚕幼虫发病率较低。通过重组质粒转染方法,可以使BmDNV-ZJ感受性品种和抵抗性品种都发病,但感受性品种的发病率较高。将重组质粒转染发病蚕的中肠匀浆,取上清液经口接种健康蚕幼虫,也能使其发病。上述结果表明,携带BmDNV-ZJ DNA的重组质粒在家蚕幼虫体内拯救出了感染性的病毒粒子。
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| 关 键 词： | 家蚕 浓核病毒 重组质粒 病毒拯救 |
Cloning of the Genome of Bombyx mori Densovirus Zhenjiang Strain and Rescue of Infectious Virions From Recombinant Plasmids in the Silkworm |
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| FU Yan-Hong,TANG Shun-Ming,QIN Guang-Xing,LIU Ting,GUO Xi-Jie.Cloning of the Genome of Bombyx mori Densovirus Zhenjiang Strain and Rescue of Infectious Virions From Recombinant Plasmids in the Silkworm[J].Acta Sericologica Sinica,2008,34(3). |
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| Authors: | FU Yan-Hong TANG Shun-Ming QIN Guang-Xing LIU Ting GUO Xi-Jie |
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| Abstract: | The genome of Bombyx mori Densovirus Zhenjiang Strain(BmDNV-ZJ) consists of two kinds of DNA molecules,VD1 and VD2.With the total DNA extracted from the midgut of BmDNV-ZJ infected silkworm larvae as template,VD1 and VD2 were amplified by polymerase chain reaction(PCR) and cloned into the plasmid pGEM-T to construct recombinant plasmids pGEM-VD1 and pGEM-VD2.With the mediation of DEAE-dextran,the recombinant plasmids were transfected into silkworm larvae of susceptible and non-susceptible varieties respectively,which resulted in the occurrence of the viral disease to the silkworm,while with lower infection rate in the non-susceptible variety.The disease was confirmed by detection of virions with immunodiffusion and immunoenzyme histochemical method.However,the transfection of pGEM-VD2 resulted in only lower morbidity to the silkworm than that of pGEM-VD1 did.The virions extracted from transfected larvae were as infectious to silkworm as wild-type ones.These results revealed that the viral genome could be rescued from the recombinant plasmids pGEM-VD1 and pGEM-VD2 by transfection of the plasmid DNA to silkworm larvae. |
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| Keywords: | Bombyx mori Densovirus Recombinant plasmid Virus rescue |
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