Efficient identification of recombinant adenoviruses by direct plaque screening

被引:15
作者
Lu, Y [1 ]
Zhang, Y [1 ]
Steiner, MS [1 ]
机构
[1] Univ Tennessee, Coll Med, Dept Urol, Urol Res Labs, Memphis, TN 38163 USA
关键词
D O I
10.1089/dna.1998.17.643
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:643 / 645
页数:3
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