Efficient recovery and regeneration of integrated retroviruses
Efficient recovery and regeneration of integrated retroviruses
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DOI：
10.1093/nar/25.14.2940
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发表时间：
1997-07-15
影响因子：
14.9
通讯作者：
Roninson, IB
中科院分区：
文献类型：
--
作者：
Schott, B;Kandel, ES;Roninson, IB
我们报告了一种快速有效的基于 PCR 的整合重组救援程序。逆转录病毒。使用一对源自长末端重复序列 (LTR) 的引物，通过长程 PCR 扩增全长原病毒 DNA，并通过用 PCR 衍生的原病毒转染逆转录病毒包装细胞来再生病毒。 PCR 产物的病毒产量与逆转录病毒质粒载体的病毒产量相似，并且不同插入片段的表达在回收的逆转录病毒群体中得到准确维持。该程序适用于从逆转录病毒文库进行表达克隆，并且应适用于天然逆转录病毒群体的分析。
We report a rapid and efficient PCR-based rescue procedure for integrated recombinant. retroviruses. Full-length proviral DNA is amplified by long-range PCR using a pair of primers derived from the long terminal repeats (LTR), and virus is regenerated by transfecting retrovirus-packaging cells with the PCR-derived provirus. The viral yield from the PCR product is similar to that from the retroviral plasmid vector, and the representation of different inserts is accurately maintained in the recovered retroviral population. This procedure is suitable for expression cloning from retroviral libraries and should be applicable to the analysis of natural retrovirus populations.