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Open Access Highly Accessed Methodology article

Flip-Flop HSV-BAC: bacterial artificial chromosome based system for rapid generation of recombinant herpes simplex virus vectors using two independent site-specific recombinases

Toshihiko Kuroda1*, Robert L Martuza1, Tomoki Todo12 and Samuel D Rabkin1

Author Affiliations

1 Molecular Neurosurgery Laboratory, Department of Neurosurgery, Massachusetts General Hospital/Harvard Medical School, 185 Cambridge St., CPZN-3800, Boston, MA 02114, USA

2 Present address: Department of Neurosurgery, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113–8655, Japan

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BMC Biotechnology 2006, 6:40  doi:10.1186/1472-6750-6-40

Published: 22 September 2006

Additional files

Additional File 1:

Genomic structure of M24-BAC and bM24-BAC viral DNA. HindIII restriction analysis of purified viral DNAs obtained from M24-BAC and bM24-BAC virus isolates #1 and #2, which are identical.

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Additional File 2:

Replication of M24-BAC and bM24-BAC in E5 cells. (a) Replication assay of recombinant viruses. 3.8 × 105 E5 cells grown in 12-well plates were infected with the indicated viruses at an MOI of 0.02, viruses were harvested at the indicated times and titered on E5 cells. (b) Virus yield of indicated viruses 74 hours after infection of E5 cells at an MOI of 0.02, from the same experiment as in a. (Error bars show standard deviation. N = 3)

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Additional File 3:

Schematic Hind III restriction map of the UL39 region in the recombinant viruses and BAC plasmids. The numbers in red show the lengths in kb of HindIII restriction fragments. H, HindIII.

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