A BAC clone of MDV strain GX0101 with REV-LTR integration retained its pathogenicity
A BAC clone of MDV strain GX0101 with REV-LTR integration retained its pathogenicity
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DOI：
10.1007/s11434-009-0364-3
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发表时间：
2009-08
影响因子：
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通讯作者：
Ai-jun Sun;P. Lawrence;Yuguang Zhao;Yanpeng Li;V. Nair;Z. Cui
中科院分区：
文献类型：
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作者：
Ai-jun Sun;P. Lawrence;Yuguang Zhao;Yanpeng Li;V. Nair;Z. Cui
将马立克氏病病毒(MDV)GX0101株的全基因组克隆到细菌人工染色体(BAC)中，获得了马立克氏病病毒(MDV)GX0101株的全基因组。通过同源重组将BAC载体序列导入MDV基因组的US2基因座。用含BAC载体的病毒DNA转化大肠杆菌DH10B。将重组BAC DNA转入原代鸡胚成纤维细胞(CEF)，成功地拯救了重组病毒。该BAC病毒克隆被命名为BAC-GX0101。将重组病毒接种1日龄雏鸡，最早在感染后62d即可检出内脏肿瘤。BAC来源的病毒与其亲本病毒在生长能力和对禽类的致病性上没有差异。BAC来源的病毒保持了其致瘤性和免疫抑制作用。综上所述，成功地将GX0101株的全基因组克隆到BAC中，挽救了感染性克隆。通过强大的BAC操纵系统，该感染性克隆将为进一步了解插入的REV-LTR序列在MDV GX0101株中的功能作用提供有用的工具。
The complete genome of Marek’s disease virus (MDV) strain GX0101, which was integrated with the LTR sequences of REV, was cloned inEscherichia colias a bacterial artificial chromosome (BAC). BAC vector sequences were introduced into the US2 locus of the MDV genome by homologous recombination. The viral DNA containing the BAC vector was used to transformEscherichia colistrain of DH10B. Then the recombinant virus was successfully rescued by transfection of the recombinant BAC DNA into primary chicken embryo fibroblast (CEF). This BAC viral clone was named bac-GX0101. When the reconstituted virus was inoculated into 1-day-old birds, visceral tumors could be detected as early as 62 d post infection. There was no difference in growth ability and pathogenicity to birds between the BAC derived virus and its parental virus. The BAC derived virus maintained its oncogenicity and immunosuppressive effects. In conclusion, the complete genome of GX0101 strain was successfully cloned into BAC and the infectious clone was rescued. With the powerful BAC manipulation system, the infectious clone will provide a useful tool for further understanding the functional roles of the inserted REV-LTR sequence in the GX0101 strain of MDV.