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Adenoviral vectors have shown great promise as vaccine carriers and in gene transfer to correct underlying genetic diseases. Traditionally, construction of adenoviral vectors is complex and time consuming. In this paper, we provide an improved method for efficient generation of novel adenoviral vectors by using direct cloning. We introduce a feasible and detailed protocol for the development of chimpanzee adenoviruses (Ads) as molecular clones, as well as for the generation of recombinant virus from the molecular clones. Recombinant viruses are genetically stable and induce potent immune responses in animals. Generation of new Ad molecular clones or new recombinant Ad can be achieved in 2 months or 2 weeks, respectively.

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Adeno-associated virus (AAV) is a defective, nonpathogenic human parvovirus, which coinfects with a helper adenovirus or herpes virus. AAV's unique characteristics have made it an appealing vector system for gene delivery. AAV or recombinant AAV (rAAV) has been widely detected using negative stain transmission electron microscopy (TEM) but little has been detected using atomic force microscopy (AFM). In this article, we used AFM and TEM to observe the recombinant AAV-2 (rAAV-2) virus particles and applied statistical analysis to the AFM and TEM images. The results indicated that the rAAV-2 particle was a slightly elliptic particle close to round when it was detected by TEM (the mean length of major and minor axes of rAAV-2 particles was 24.77 +/- 1.78 nm and 21.84 +/- 1.57 nm, respectively), whereas when detected by AFM, the rAAV-2 particle was almost round. Even though the dimensions of the rAAV-2 particle exhibited a polymorphous distribution via off-line particle analysis of AFM, most of the rAAV-2 particles had a mean diameter of approximate 22.04 nm, which was similar to the results obtained by TEM. The results above suggested that AFM was important for accurately determining the average dimensions and distributions of virus particles.