Lu Y, Zhang Y, Steiner M S
Department of Urology, University of Tennessee, Memphis 38163, USA.
DNA Cell Biol. 1998 Jul;17(7):643-5. doi: 10.1089/dna.1998.17.643.
With the increasing use of adenoviral vectors for gene transfer and gene therapy, it is crucial to produce specific recombinant adenoviruses more efficiently. One of the most time-consuming steps is to screen the unique recombinant adenovirus among the plaques, in which each plaque has to be amplified individually in kidney 293 cells in order to obtain enough adenoviruses for DNA extraction and subsequent identification. We have developed a fast and simple way to screen recombinant adenoviruses by direct plaque screening. The direct plaque-screening method employed DNA obtained from the viral plaque itself for PCR amplification and subsequent adenoviral recombinant identification. The time and labor involved in these steps has been significantly reduced.
随着腺病毒载体在基因转移和基因治疗中的应用日益广泛,更高效地生产特定重组腺病毒至关重要。最耗时的步骤之一是在噬斑中筛选独特的重组腺病毒,其中每个噬斑都必须在肾293细胞中单独扩增,以获得足够的腺病毒用于DNA提取和后续鉴定。我们开发了一种通过直接噬斑筛选来筛选重组腺病毒的快速简便方法。直接噬斑筛选方法利用从病毒噬斑本身获得的DNA进行PCR扩增及随后的腺病毒重组鉴定。这些步骤所涉及的时间和人力已大幅减少。