DOI: 10.1073/pnas.95.5.2509. A simplified system for generating recombinant adenoviruses

A simplified system for generating recombinant adenoviruses

10.1073/pnas.95.5.2509

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

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.

Bibliography

He, T.-C., Zhou, S., da Costa, L. T., Yu, J., Kinzler, K. W., & Vogelstein, B. (1998). A simplified system for generating recombinant adenoviruses. Proceedings of the National Academy of Sciences, 95(5), 2509â2514.

Authors 6

Tong-Chuan He (first)
Shibin Zhou (additional)
Luis T. da Costa (additional)
Jian Yu (additional)
Kenneth W. Kinzler (additional)
Bert Vogelstein (additional)

References 26 Referenced 2,996

10.1038/357455a0
10.1146/annurev.bi.62.070193.001203
F L Graham, L Prevec Methods Mol Biol 7, 109–128 (1991). / Methods Mol Biol by Graham F L (1991)
K L Berkner BioTechniques 6, 616–629 (1988). / BioTechniques by Berkner K L (1988)
T Shenk Fields Virology, eds B N Fields, D M Knipe, P M Howley, R M Chanock, J L Melnick, T P Monath, B Roizman, S E Straus (Lippincott, Philadelphia), pp. 2111–2148 (1996). / Fields Virology by Shenk T (1996)
M S Horwitz Fields Virology, eds B N Fields, D M Knipe, P M Howley, R M Chanock, J L Melnick, T P Monath, B Roizman, S E Straus (Lippincott, Philadelphia), pp. 2149–2171 (1996). / Fields Virology by Horwitz M S (1996)
10.1002/j.1460-2075.1985.tb04158.x
10.1126/science.2017680
10.1016/0168-1702(93)90090-A
10.1172/JCI115902
10.1099/0022-1317-36-1-59
10.1016/S0091-679X(08)60603-2
10.1073/pnas.91.13.6186
10.1128/jvi.70.7.4805-4810.1996
10.1073/pnas.94.4.1414
10.1016/0378-1119(92)90691-H
10.1089/hum.1996.7.2-215
10.1016/S0022-2836(83)80284-8
10.1128/mcb.14.2.1333-1346.1994
10.1016/S0091-679X(08)60603-2
10.1016/0092-8674(94)90168-6
10.1146/annurev.ge.29.120195.002453
10.1093/nar/11.17.6003
10.1128/mcb.4.2.302-309.1984
10.1016/S1097-2765(00)80002-7
10.1128/jvi.71.3.1842-1849.1997

Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 10:35 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 4:39 p.m.)
Indexed	1 week, 1 day ago (Aug. 21, 2025, 12:44 p.m.)
Issued	27 years, 5 months ago (March 3, 1998)
Published	27 years, 5 months ago (March 3, 1998)
Published Online	27 years, 5 months ago (March 3, 1998)
Published Print	27 years, 5 months ago (March 3, 1998)

Funders 0

None

BibTeX

@article{He_1998, title={A simplified system for generating recombinant adenoviruses}, volume={95}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.95.5.2509}, DOI={10.1073/pnas.95.5.2509}, number={5}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={He, Tong-Chuan and Zhou, Shibin and da Costa, Luis T. and Yu, Jian and Kinzler, Kenneth W. and Vogelstein, Bert}, year={1998}, month=mar, pages={2509–2514} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        8,
        21
      ]
    ],
    "date-time": "2025-08-21T16:44:25Z",
    "timestamp": 1755794665755
  },
  "reference-count": 26,
  "publisher": "Proceedings of the National Academy of Sciences",
  "issue": "5",
  "content-domain": {
    "domain": [
      "www.pnas.org"
    ],
    "crossmark-restriction": true
  },
  "published-print": {
    "date-parts": [
      [
        1998,
        3,
        3
      ]
    ]
  },
  "abstract": "<jats:p>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.</jats:p>",
  "DOI": "10.1073/pnas.95.5.2509",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2002,
        7,
        26
      ]
    ],
    "date-time": "2002-07-26T14:35:50Z",
    "timestamp": 1027694150000
  },
  "page": "2509-2514",
  "update-policy": "http://dx.doi.org/10.1073/pnas.cm10313",
  "source": "Crossref",
  "is-referenced-by-count": 2996,
  "title": "A simplified system for generating recombinant adenoviruses",
  "prefix": "10.1073",
  "volume": "95",
  "author": [
    {
      "given": "Tong-Chuan",
      "family": "He",
      "sequence": "first",
      "affiliation": [
        {
          "name": "The Howard Hughes Medical Institute, The Program in Human Genetics and Molecular Biology, The Johns Hopkins Oncology Center, 424 North Bond Street, Baltimore, MD 21231"
        }
      ]
    },
    {
      "given": "Shibin",
      "family": "Zhou",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "The Howard Hughes Medical Institute, The Program in Human Genetics and Molecular Biology, The Johns Hopkins Oncology Center, 424 North Bond Street, Baltimore, MD 21231"
        }
      ]
    },
    {
      "given": "Luis T.",
      "family": "da Costa",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "The Howard Hughes Medical Institute, The Program in Human Genetics and Molecular Biology, The Johns Hopkins Oncology Center, 424 North Bond Street, Baltimore, MD 21231"
        }
      ]
    },
    {
      "given": "Jian",
      "family": "Yu",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "The Howard Hughes Medical Institute, The Program in Human Genetics and Molecular Biology, The Johns Hopkins Oncology Center, 424 North Bond Street, Baltimore, MD 21231"
        }
      ]
    },
    {
      "given": "Kenneth W.",
      "family": "Kinzler",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "The Howard Hughes Medical Institute, The Program in Human Genetics and Molecular Biology, The Johns Hopkins Oncology Center, 424 North Bond Street, Baltimore, MD 21231"
        }
      ]
    },
    {
      "given": "Bert",
      "family": "Vogelstein",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "The Howard Hughes Medical Institute, The Program in Human Genetics and Molecular Biology, The Johns Hopkins Oncology Center, 424 North Bond Street, Baltimore, MD 21231"
        }
      ]
    }
  ],
  "member": "341",
  "published-online": {
    "date-parts": [
      [
        1998,
        3,
        3
      ]
    ]
  },
  "reference": [
    {
      "key": "e_1_3_3_1_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/357455a0"
    },
    {
      "key": "e_1_3_3_2_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1146/annurev.bi.62.070193.001203"
    },
    {
      "key": "e_1_3_3_3_2",
      "first-page": "109",
      "volume": "7",
      "author": "Graham F L",
      "year": "1991",
      "unstructured": "F L Graham, L Prevec Methods Mol Biol 7, 109\u2013128 (1991).",
      "journal-title": "Methods Mol Biol"
    },
    {
      "key": "e_1_3_3_4_2",
      "first-page": "616",
      "volume": "6",
      "author": "Berkner K L",
      "year": "1988",
      "unstructured": "K L Berkner BioTechniques 6, 616\u2013629 (1988).",
      "journal-title": "BioTechniques"
    },
    {
      "key": "e_1_3_3_5_2",
      "first-page": "2111",
      "volume-title": "Fields Virology",
      "author": "Shenk T",
      "year": "1996",
      "unstructured": "T Shenk Fields Virology, eds B N Fields, D M Knipe, P M Howley, R M Chanock, J L Melnick, T P Monath, B Roizman, S E Straus (Lippincott, Philadelphia), pp. 2111\u20132148 (1996)."
    },
    {
      "key": "e_1_3_3_6_2",
      "first-page": "2149",
      "volume-title": "Fields Virology",
      "author": "Horwitz M S",
      "year": "1996",
      "unstructured": "M S Horwitz Fields Virology, eds B N Fields, D M Knipe, P M Howley, R M Chanock, J L Melnick, T P Monath, B Roizman, S E Straus (Lippincott, Philadelphia), pp. 2149\u20132171 (1996)."
    },
    {
      "key": "e_1_3_3_7_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/j.1460-2075.1985.tb04158.x"
    },
    {
      "key": "e_1_3_3_8_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.2017680"
    },
    {
      "key": "e_1_3_3_9_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0168-1702(93)90090-A"
    },
    {
      "key": "e_1_3_3_10_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1172/JCI115902"
    },
    {
      "key": "e_1_3_3_11_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1099/0022-1317-36-1-59"
    },
    {
      "key": "e_1_3_3_12_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0091-679X(08)60603-2"
    },
    {
      "key": "e_1_3_3_13_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.91.13.6186"
    },
    {
      "key": "e_1_3_3_14_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jvi.70.7.4805-4810.1996"
    },
    {
      "key": "e_1_3_3_15_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.94.4.1414"
    },
    {
      "key": "e_1_3_3_16_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0378-1119(92)90691-H"
    },
    {
      "key": "e_1_3_3_17_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1089/hum.1996.7.2-215"
    },
    {
      "key": "e_1_3_3_18_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0022-2836(83)80284-8"
    },
    {
      "key": "e_1_3_3_19_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/mcb.14.2.1333-1346.1994"
    },
    {
      "key": "e_1_3_3_20_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0091-679X(08)60603-2"
    },
    {
      "key": "e_1_3_3_21_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(94)90168-6"
    },
    {
      "key": "e_1_3_3_22_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1146/annurev.ge.29.120195.002453"
    },
    {
      "key": "e_1_3_3_23_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/nar/11.17.6003"
    },
    {
      "key": "e_1_3_3_24_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/mcb.4.2.302-309.1984"
    },
    {
      "key": "e_1_3_3_25_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S1097-2765(00)80002-7"
    },
    {
      "key": "e_1_3_3_26_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jvi.71.3.1842-1849.1997"
    }
  ],
  "container-title": "Proceedings of the National Academy of Sciences",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://pnas.org/doi/pdf/10.1073/pnas.95.5.2509",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2022,
        4,
        13
      ]
    ],
    "date-time": "2022-04-13T20:39:07Z",
    "timestamp": 1649882347000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://pnas.org/doi/full/10.1073/pnas.95.5.2509"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        1998,
        3,
        3
      ]
    ]
  },
  "references-count": 26,
  "journal-issue": {
    "issue": "5",
    "published-print": {
      "date-parts": [
        [
          1998,
          3,
          3
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1073/pnas.95.5.2509"
  ],
  "URL": "http://dx.doi.org/10.1073/pnas.95.5.2509",
  "relation": {},
  "ISSN": [
    "0027-8424",
    "1091-6490"
  ],
  "subject": [],
  "container-title-short": "Proc. Natl. Acad. Sci. U.S.A.",
  "published": {
    "date-parts": [
      [
        1998,
        3,
        3
      ]
    ]
  },
  "assertion": [
    {
      "value": "1997-12-30",
      "order": 1,
      "name": "accepted",
      "label": "Accepted",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    },
    {
      "value": "1998-03-03",
      "order": 2,
      "name": "published",
      "label": "Published",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    }
  ]
}