By T.R. Flotte, and K.I. Berns (Eds.)
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Additional resources for Adeno-Associated Viral Vectors for Gene Therapy
The NaCl concentration of the peak is adjusted to 1 M and subjected to hydrophobic interaction chromatography on Phenyl sepharose (5 ml column, Pharmacia); the flow‐through is collected, Fig. 3B. The flow‐through is diluted to ~150 mM NaCl by diluting with water. 5 M), Fig. 3C. Absorbance is monitored at 280 nm, the peak is collected and the presence of virus is confirmed in peak fractions by infectious titer, vector genome titer, and silver staining of SDS‐PAGE gels. In a typical large‐scale run, the vector genome titers (DNase resistant particles) average 2–3 Â 1013 ml–1 in a total of 10 ml.
2003). Genetic fate of recombinant adeno‐associated virus vector genomes in muscle. J. Virol. 77, 3495–3504. , Chang, L. S. and Shenk, T. (1991). Transcriptional repression by YY1, a human GLI‐Kruppel‐related protein, and relief of repression by adenovirus E1A protein. Cell 67, 377–388. Smith, R. H. and Kotin, R. M. (1998). The Rep52 gene product of adeno‐ associated virus is a DNA helicase with 30 ‐to–50 polarity. J. Virol. 72, 4874–4881. Ch. 2 PRODUCTION OF RESEARCH AND CLINICAL AAV 55 Smith, R.
Smith, R. H. and Kotin, R. M. (1998). The Rep52 gene product of adeno‐ associated virus is a DNA helicase with 30 ‐to–50 polarity. J. Virol. 72, 4874–4881. Ch. 2 PRODUCTION OF RESEARCH AND CLINICAL AAV 55 Smith, R. , Spano, A. J. and Kotin, R. M. (1997). The Rep78 gene product of adeno‐associated virus (AAV) self‐associates to form a hexameric complex in the presence of AAV ori sequences. J. Virol. 71, 4461–4471. Snyder, R. , Xiao, X. and Samulski, R. J. (1996). Production of recombinant adeno‐associated viral vectors.
Adeno-Associated Viral Vectors for Gene Therapy by T.R. Flotte, and K.I. Berns (Eds.)