A simplified system for generating recombinant adenoviruses.
Recombinant adenoviruses provide a versatile system for gene expression studies and therapeutic applications. We report herein a strategy that simplifies the generation and production of such viruses. A recombinant adenoviral plasmid is generated with a minimum of enzymatic manipulations, using homologous recombination in bacteria rather than in eukaryotic cells. After transfections of such plasmids into a mammalian packaging cell line, viral production is conveniently followed with the aid of green fluorescent protein, encoded by a gene incorporated into the viral backbone. Homogeneous viruses can be obtained from this procedure without plaque purification. This system should expedite the process of generating and testing recombinant adenoviruses for a variety of purposes.
He
T C
TC
The Howard Hughes Medical Institute, 424 North Bond Street, Baltimore, MD 21231, USA. tche@welchlink.welch.jhu.edu
Zhou
S
S
da Costa
L T
LT
Yu
J
J
Kinzler
K W
KW
Vogelstein
B
B
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002460:Cell Line; D004926:Escherichia coli / Q000235:genetics; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006801:Humans; D007668:Kidney; D008164:Luminescent Proteins / Q000096:biosynthesis; D008322:Mammals; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D011995:Recombination, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D014162:Transfection / Q000379:methods*
D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins