ZHANG Xiao-Dong, ZHANG Jia-Min, GUO Hai-Chao, SHU Li-Hua and HU Yuan-Yang. Rescue of Infectious Virions from Recom binant Plasmid with the Genome of Periplanta liginosa,Densovirus[J]. Virologica Sinica, 1999, 14(2): 152-156.
Citation:
ZHANG Xiao-Dong, ZHANG Jia-Min, GUO Hai-Chao, SHU Li-Hua, HU Yuan-Yang.
Rescue of Infectious Virions from Recom binant Plasmid with the Genome of Periplanta liginosa,Densovirus .VIROLOGICA SINICA, 1999, 14(2)
: 152-156.
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摘要
用Glassmilk法纯化PfDNVdsDNA,在其3端和PstⅠ切开的pUC119质粒的3端分别加dG和dC尾,连接、转化、筛选得到分子量为8.7kb的重组质粒。通过酶切证明插入DNA为PfDNV全基因组DNA。利用DEAEdextran转染技术,将此重组质粒导入黑胸大蠊幼虫体内,此重组质粒能使虫体发病死亡。电镜超薄切片观察发现虫体细胞内存在大量的病毒粒子。同样,在免疫扩散实验中虫体PBS浸出液能与抗PfDNV的抗体产生沉淀线。用重组质粒感染致死的虫体喂食健康黑胸大蠊,也能使其发病死亡,通过电镜可以观察到在细胞内增殖的病毒粒子。
Rescue of Infectious Virions from Recom binant Plasmid with the Genome of Periplanta liginosa,Densovirus
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Abstract
An infectious genome of the Periplanta fuliginosa densovirus (PfDNV) has been cloned into the bacterial plasmid pUC119. The viral genome could be rescued from the recombinant plasmid PfNDV pUC119 by transfection to sensitive Periplanta fuliginosa larvae. Virions can be found in the cells of the transfected larvaes by electron microscope, and the rescued virions were as infectious as wild type virions. In contrast, the viral genome could not be rescued from the recombinant plasmid deleted one extremity.
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References
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Proportional views
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