DOI: 10.1126/science.233.4762.478. Engineering Herbicide Tolerance in Transgenic Plants

Engineering Herbicide Tolerance in Transgenic Plants

10.1126/science.233.4762.478

Crossref journal-article

American Association for the Advancement of Science (AAAS)

Science (221)

Abstract

The herbicide glyphosate is a potent inhibitor of the enzyme 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase in higher plants. A complementary DNA (cDNA) clone encoding EPSP synthase was isolated from a complementary DNA library of a glyphosate-tolerant Petunia hybrida cell line (MP4-G) that overproduces the enzyme. This cell line was shown to overproduce EPSP synthase messenger RNA as a result of a 20-fold amplification of the gene. A chimeric EPSP synthase gene was constructed with the use of the cauliflower mosaic virus 35 S promoter to attain high level expression of EPSP synthase and introduced into petunia cells. Transformed petunia cells as well as regenerated transgenic plants were tolerant to glyphosate.

Bibliography

Shah, D. M., Horsch, R. B., Klee, H. J., Kishore, G. M., Winter, J. A., Tumer, N. E., Hironaka, C. M., Sanders, P. R., Gasser, C. S., Aykent, S., Siegel, N. R., Rogers, S. G., & Fraley, R. T. (1986). Engineering Herbicide Tolerance in Transgenic Plants. Science, 233(4762), 478â481.

Authors 13

Dilip M. Shah (first)
Robert B. Horsch (additional)
Harry J. Klee (additional)
Ganesh M. Kishore (additional)
Jill A. Winter (additional)
Nilgun E. Tumer (additional)
Cathy M. Hironaka (additional)
Patricia R. Sanders (additional)
Charles S. Gasser (additional)
Serdar Aykent (additional)
Ned R. Siegel (additional)
Stephen G. Rogers (additional)
Robert T. Fraley (additional)

References 24 Referenced 403

AMRHEIN, N, BIOCHEMICAL BASIS FOR GLYCOPHOSPHATE-TOLERANCE IN A BACTERIUM AND A PLANT-TISSUE CULTURE, FEBS LETTERS 157: 191 (1983). (10.1016/0014-5793(83)81143-0) / FEBS LETTERS (1983)
COMAI, L, EXPRESSION IN PLANTS OF A MUTANT AROA GENE FROM SALMONELLA-TYPHIMURIUM CONFERS TOLERANCE TO GLYPHOSATE, NATURE 317: 741 (1985). (10.1038/317741a0) / NATURE (1985)
DELLACIOPPA G COMMUNICATION.
FEDOROFF, N, JOURNAL OF MOLECULAR AND APPLIED GENETICS 2: 11 (1983). / JOURNAL OF MOLECULAR AND APPLIED GENETICS (1983)
FRALEY, R.T., THE SEV SYSTEM - A NEW DISARMED TI-PLASMID VECTOR SYSTEM FOR PLANT TRANSFORMATION, BIO-TECHNOLOGY 3: 629 (1985). / BIO-TECHNOLOGY (1985)
10.1126/science.227.4691.1229
HUNKAPILLER, M.W., ISOLATION OF MICROGRAM QUANTITIES OF PROTEINS FROM POLYACRYLAMIDE GELS FOR AMINO-ACID SEQUENCE-ANALYSIS, METHODS IN ENZYMOLOGY 91: 227 (1983). (10.1016/S0076-6879(83)91019-4) / METHODS IN ENZYMOLOGY (1983)
HUNKAPILLER, M.W., HIGH-SENSITIVITY SEQUENCING WITH A GAS-PHASE SEQUENATOR, METHODS IN ENZYMOLOGY 91: 399 (1983). (10.1016/S0076-6879(83)91038-8) / METHODS IN ENZYMOLOGY (1983)
HUYNH, T.V., DNA CLONING PRACTICA: 49 (1985). / DNA CLONING PRACTICA (1985)
KLEE H unpublished data.
Maniatis T. Molecular Cloning: A Laboratory Manual (1982).
MOUSDALE, D.M., PURIFICATION AND PROPERTIES OF 5-ENOLPYRUVYLSHIKIMATE 3-PHOSPHATE SYNTHASE FROM SEEDLINGS OF PISUM-SATIVUM-L, PLANTA 160: 78 (1984). (10.1007/BF00392469) / PLANTA (1984)
MOUSDALE, D.M., SUBCELLULAR-LOCALIZATION OF THE COMMON SHIKIMATE-PATHWAY ENZYMES IN PISUM-SATIVUM-L, PLANTA 163: 241 (1985). (10.1007/BF00393514) / PLANTA (1985)
NAFZIGER, E.D., SELECTION AND CHARACTERIZATION OF A CARROT CELL-LINE TOLERANT TO GLYPHOSATE, PLANT PHYSIOLOGY 76: 571 (1984). (10.1104/pp.76.3.571) / PLANT PHYSIOLOGY (1984)
ODELL, J.T., IDENTIFICATION OF DNA-SEQUENCES REQUIRED FOR ACTIVITY OF THE CAULIFLOWER MOSAIC VIRUS-35S PROMOTER, NATURE 313: 810 (1985). (10.1038/313810a0) / NATURE (1985)
RUBIN, J.L., GLYPHOSATE INHIBITION OF 5-ENOLPYRUVYLSHIKIMATE 3-PHOSPHATE SYNTHASE FROM SUSPENSION-CULTURED CELLS OF NICOTIANA-SILVESTRIS, PLANT PHYSIOLOGY 75: 839 (1984). (10.1104/pp.75.3.839) / PLANT PHYSIOLOGY (1984)
SCHELLER, R.H., A FAMILY OF GENES THAT CODES FOR ELH, A NEUROPEPTIDE ELICITING A STEREOTYPED PATTERN OF BEHAVIOR IN APLYSIA, CELL 28: 707 (1982). (10.1016/0092-8674(82)90050-2) / CELL (1982)
SHAH D.M. unpublished data.
SHURE, M, MOLECULAR-IDENTIFICATION AND ISOLATION OF THE WAXY LOCUS IN MAIZE, CELL 35: 225 (1983). (10.1016/0092-8674(83)90225-8) / CELL (1983)
SMART, C.C., SELECTIVE OVERPRODUCTION OF 5-ENOL-PYRUVYLSHIKIMIC ACID 3-PHOSPHATE SYNTHASE IN A PLANT-CELL CULTURE WHICH TOLERATES HIGH-DOSES OF THE HERBICIDE GLYPHOSATE, JOURNAL OF BIOLOGICAL CHEMISTRY 260: 16338 (1985). (10.1016/S0021-9258(17)36242-7) / JOURNAL OF BIOLOGICAL CHEMISTRY (1985)
STEINRUCKEN, H.C., OVERPRODUCTION OF 5-ENOLPYRUVYLSHIKIMATE-3-PHOSPHATE SYNTHASE IN A GLYPHOSATE-TOLERANT PETUNIA-HYBRIDA CELL-LINE, ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS 244: 169 (1986). (10.1016/0003-9861(86)90106-2) / ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS (1986)
STEINRUCKEN, H.C., THE HERBICIDE GLYPHOSATE IS A POTENT INHIBITOR OF 5-ENOLPYRUVYL-SHIKIMIC-ACID 3-PHOSPHATE SYNTHASE, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 94: 1207 (1980). (10.1016/0006-291X(80)90547-1) / BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS (1980)
SUGGS, S, ICN UCLA S DEV BIOL: 683 (1981). / ICN UCLA S DEV BIOL (1981)
ZOLLER, M.J., OLIGONUCLEOTIDE-DIRECTED MUTAGENESIS OF DNA FRAGMENTS CLONED INTO M13 VECTORS, METHODS IN ENZYMOLOGY 100: 468 (1983). (10.1016/0076-6879(83)00074-9) / METHODS IN ENZYMOLOGY (1983)

Dates

Type	When
Created	18 years, 11 months ago (Oct. 5, 2006, 4:28 p.m.)
Deposited	1 year, 7 months ago (Jan. 12, 2024, 11 a.m.)
Indexed	1 month ago (Aug. 7, 2025, 4:55 a.m.)
Issued	39 years, 1 month ago (July 25, 1986)
Published	39 years, 1 month ago (July 25, 1986)
Published Print	39 years, 1 month ago (July 25, 1986)

Funders 0

None

BibTeX

@article{Shah_1986, title={Engineering Herbicide Tolerance in Transgenic Plants}, volume={233}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.233.4762.478}, DOI={10.1126/science.233.4762.478}, number={4762}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Shah, Dilip M. and Horsch, Robert B. and Klee, Harry J. and Kishore, Ganesh M. and Winter, Jill A. and Tumer, Nilgun E. and Hironaka, Cathy M. and Sanders, Patricia R. and Gasser, Charles S. and Aykent, Serdar and Siegel, Ned R. and Rogers, Stephen G. and Fraley, Robert T.}, year={1986}, month=jul, pages={478–481} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        8,
        7
      ]
    ],
    "date-time": "2025-08-07T08:55:35Z",
    "timestamp": 1754556935097
  },
  "reference-count": 24,
  "publisher": "American Association for the Advancement of Science (AAAS)",
  "issue": "4762",
  "content-domain": {
    "domain": [],
    "crossmark-restriction": false
  },
  "published-print": {
    "date-parts": [
      [
        1986,
        7,
        25
      ]
    ]
  },
  "abstract": "<jats:p>\n            The herbicide glyphosate is a potent inhibitor of the enzyme 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase in higher plants. A complementary DNA (cDNA) clone encoding EPSP synthase was isolated from a complementary DNA library of a glyphosate-tolerant\n            <jats:italic>Petunia hybrida</jats:italic>\n            cell line (MP4-G) that overproduces the enzyme. This cell line was shown to overproduce EPSP synthase messenger RNA as a result of a 20-fold amplification of the gene. A chimeric EPSP synthase gene was constructed with the use of the cauliflower mosaic virus 35\n            <jats:italic>S</jats:italic>\n            promoter to attain high level expression of EPSP synthase and introduced into petunia cells. Transformed petunia cells as well as regenerated transgenic plants were tolerant to glyphosate.\n          </jats:p>",
  "DOI": "10.1126/science.233.4762.478",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2006,
        10,
        5
      ]
    ],
    "date-time": "2006-10-05T20:28:52Z",
    "timestamp": 1160080132000
  },
  "page": "478-481",
  "source": "Crossref",
  "is-referenced-by-count": 403,
  "title": "Engineering Herbicide Tolerance in Transgenic Plants",
  "prefix": "10.1126",
  "volume": "233",
  "author": [
    {
      "given": "Dilip M.",
      "family": "Shah",
      "sequence": "first",
      "affiliation": [
        {
          "name": "Plant Molecular Biology, Division of Biological Sciences, Monsanto Company, 700 Chesterfield Village Parkway, St. Louis, MO 63198."
        }
      ]
    },
    {
      "given": "Robert B.",
      "family": "Horsch",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Plant Molecular Biology, Division of Biological Sciences, Monsanto Company, 700 Chesterfield Village Parkway, St. Louis, MO 63198."
        }
      ]
    },
    {
      "given": "Harry J.",
      "family": "Klee",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Plant Molecular Biology, Division of Biological Sciences, Monsanto Company, 700 Chesterfield Village Parkway, St. Louis, MO 63198."
        }
      ]
    },
    {
      "given": "Ganesh M.",
      "family": "Kishore",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Plant Molecular Biology, Division of Biological Sciences, Monsanto Company, 700 Chesterfield Village Parkway, St. Louis, MO 63198."
        }
      ]
    },
    {
      "given": "Jill A.",
      "family": "Winter",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Plant Molecular Biology, Division of Biological Sciences, Monsanto Company, 700 Chesterfield Village Parkway, St. Louis, MO 63198."
        }
      ]
    },
    {
      "given": "Nilgun E.",
      "family": "Tumer",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Plant Molecular Biology, Division of Biological Sciences, Monsanto Company, 700 Chesterfield Village Parkway, St. Louis, MO 63198."
        }
      ]
    },
    {
      "given": "Cathy M.",
      "family": "Hironaka",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Plant Molecular Biology, Division of Biological Sciences, Monsanto Company, 700 Chesterfield Village Parkway, St. Louis, MO 63198."
        }
      ]
    },
    {
      "given": "Patricia R.",
      "family": "Sanders",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Plant Molecular Biology, Division of Biological Sciences, Monsanto Company, 700 Chesterfield Village Parkway, St. Louis, MO 63198."
        }
      ]
    },
    {
      "given": "Charles S.",
      "family": "Gasser",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Plant Molecular Biology, Division of Biological Sciences, Monsanto Company, 700 Chesterfield Village Parkway, St. Louis, MO 63198."
        }
      ]
    },
    {
      "given": "Serdar",
      "family": "Aykent",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Plant Molecular Biology, Division of Biological Sciences, Monsanto Company, 700 Chesterfield Village Parkway, St. Louis, MO 63198."
        }
      ]
    },
    {
      "given": "Ned R.",
      "family": "Siegel",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Plant Molecular Biology, Division of Biological Sciences, Monsanto Company, 700 Chesterfield Village Parkway, St. Louis, MO 63198."
        }
      ]
    },
    {
      "given": "Stephen G.",
      "family": "Rogers",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Plant Molecular Biology, Division of Biological Sciences, Monsanto Company, 700 Chesterfield Village Parkway, St. Louis, MO 63198."
        }
      ]
    },
    {
      "given": "Robert T.",
      "family": "Fraley",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Plant Molecular Biology, Division of Biological Sciences, Monsanto Company, 700 Chesterfield Village Parkway, St. Louis, MO 63198."
        }
      ]
    }
  ],
  "member": "221",
  "reference": [
    {
      "key": "e_1_2_1_1_1",
      "doi-asserted-by": "crossref",
      "first-page": "191",
      "DOI": "10.1016/0014-5793(83)81143-0",
      "volume": "157",
      "year": "1983",
      "unstructured": "AMRHEIN, N, BIOCHEMICAL BASIS FOR GLYCOPHOSPHATE-TOLERANCE IN A BACTERIUM AND A PLANT-TISSUE CULTURE, FEBS LETTERS 157: 191 (1983).",
      "journal-title": "FEBS LETTERS"
    },
    {
      "key": "e_1_2_1_2_1",
      "doi-asserted-by": "crossref",
      "first-page": "741",
      "DOI": "10.1038/317741a0",
      "volume": "317",
      "year": "1985",
      "unstructured": "COMAI, L, EXPRESSION IN PLANTS OF A MUTANT AROA GENE FROM SALMONELLA-TYPHIMURIUM CONFERS TOLERANCE TO GLYPHOSATE, NATURE 317: 741 (1985).",
      "journal-title": "NATURE"
    },
    {
      "key": "e_1_2_1_3_1",
      "unstructured": "DELLACIOPPA G COMMUNICATION."
    },
    {
      "key": "e_1_2_1_4_1",
      "first-page": "11",
      "volume": "2",
      "year": "1983",
      "unstructured": "FEDOROFF, N, JOURNAL OF MOLECULAR AND APPLIED GENETICS 2: 11 (1983).",
      "journal-title": "JOURNAL OF MOLECULAR AND APPLIED GENETICS"
    },
    {
      "key": "e_1_2_1_5_1",
      "first-page": "629",
      "volume": "3",
      "year": "1985",
      "unstructured": "FRALEY, R.T., THE SEV SYSTEM - A NEW DISARMED TI-PLASMID VECTOR SYSTEM FOR PLANT TRANSFORMATION, BIO-TECHNOLOGY 3: 629 (1985).",
      "journal-title": "BIO-TECHNOLOGY"
    },
    {
      "key": "e_1_2_1_6_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.227.4691.1229"
    },
    {
      "key": "e_1_2_1_7_1",
      "doi-asserted-by": "crossref",
      "first-page": "227",
      "DOI": "10.1016/S0076-6879(83)91019-4",
      "volume": "91",
      "year": "1983",
      "unstructured": "HUNKAPILLER, M.W., ISOLATION OF MICROGRAM QUANTITIES OF PROTEINS FROM POLYACRYLAMIDE GELS FOR AMINO-ACID SEQUENCE-ANALYSIS, METHODS IN ENZYMOLOGY 91: 227 (1983).",
      "journal-title": "METHODS IN ENZYMOLOGY"
    },
    {
      "key": "e_1_2_1_8_1",
      "doi-asserted-by": "crossref",
      "first-page": "399",
      "DOI": "10.1016/S0076-6879(83)91038-8",
      "volume": "91",
      "year": "1983",
      "unstructured": "HUNKAPILLER, M.W., HIGH-SENSITIVITY SEQUENCING WITH A GAS-PHASE SEQUENATOR, METHODS IN ENZYMOLOGY 91: 399 (1983).",
      "journal-title": "METHODS IN ENZYMOLOGY"
    },
    {
      "key": "e_1_2_1_9_1",
      "first-page": "49",
      "year": "1985",
      "unstructured": "HUYNH, T.V., DNA CLONING PRACTICA: 49 (1985).",
      "journal-title": "DNA CLONING PRACTICA"
    },
    {
      "key": "e_1_2_1_10_1",
      "unstructured": "KLEE H unpublished data."
    },
    {
      "key": "e_1_2_1_11_1",
      "unstructured": "Maniatis T. Molecular Cloning: A Laboratory Manual (1982)."
    },
    {
      "key": "e_1_2_1_12_1",
      "doi-asserted-by": "crossref",
      "first-page": "78",
      "DOI": "10.1007/BF00392469",
      "volume": "160",
      "year": "1984",
      "unstructured": "MOUSDALE, D.M., PURIFICATION AND PROPERTIES OF 5-ENOLPYRUVYLSHIKIMATE 3-PHOSPHATE SYNTHASE FROM SEEDLINGS OF PISUM-SATIVUM-L, PLANTA 160: 78 (1984).",
      "journal-title": "PLANTA"
    },
    {
      "key": "e_1_2_1_13_1",
      "doi-asserted-by": "crossref",
      "first-page": "241",
      "DOI": "10.1007/BF00393514",
      "volume": "163",
      "year": "1985",
      "unstructured": "MOUSDALE, D.M., SUBCELLULAR-LOCALIZATION OF THE COMMON SHIKIMATE-PATHWAY ENZYMES IN PISUM-SATIVUM-L, PLANTA 163: 241 (1985).",
      "journal-title": "PLANTA"
    },
    {
      "key": "e_1_2_1_14_1",
      "doi-asserted-by": "crossref",
      "first-page": "571",
      "DOI": "10.1104/pp.76.3.571",
      "volume": "76",
      "year": "1984",
      "unstructured": "NAFZIGER, E.D., SELECTION AND CHARACTERIZATION OF A CARROT CELL-LINE TOLERANT TO GLYPHOSATE, PLANT PHYSIOLOGY 76: 571 (1984).",
      "journal-title": "PLANT PHYSIOLOGY"
    },
    {
      "key": "e_1_2_1_15_1",
      "doi-asserted-by": "crossref",
      "first-page": "810",
      "DOI": "10.1038/313810a0",
      "volume": "313",
      "year": "1985",
      "unstructured": "ODELL, J.T., IDENTIFICATION OF DNA-SEQUENCES REQUIRED FOR ACTIVITY OF THE CAULIFLOWER MOSAIC VIRUS-35S PROMOTER, NATURE 313: 810 (1985).",
      "journal-title": "NATURE"
    },
    {
      "key": "e_1_2_1_16_1",
      "doi-asserted-by": "crossref",
      "first-page": "839",
      "DOI": "10.1104/pp.75.3.839",
      "volume": "75",
      "year": "1984",
      "unstructured": "RUBIN, J.L., GLYPHOSATE INHIBITION OF 5-ENOLPYRUVYLSHIKIMATE 3-PHOSPHATE SYNTHASE FROM SUSPENSION-CULTURED CELLS OF NICOTIANA-SILVESTRIS, PLANT PHYSIOLOGY 75: 839 (1984).",
      "journal-title": "PLANT PHYSIOLOGY"
    },
    {
      "key": "e_1_2_1_17_1",
      "doi-asserted-by": "crossref",
      "first-page": "707",
      "DOI": "10.1016/0092-8674(82)90050-2",
      "volume": "28",
      "year": "1982",
      "unstructured": "SCHELLER, R.H., A FAMILY OF GENES THAT CODES FOR ELH, A NEUROPEPTIDE ELICITING A STEREOTYPED PATTERN OF BEHAVIOR IN APLYSIA, CELL 28: 707 (1982).",
      "journal-title": "CELL"
    },
    {
      "key": "e_1_2_1_18_1",
      "unstructured": "SHAH D.M. unpublished data."
    },
    {
      "key": "e_1_2_1_19_1",
      "doi-asserted-by": "crossref",
      "first-page": "225",
      "DOI": "10.1016/0092-8674(83)90225-8",
      "volume": "35",
      "year": "1983",
      "unstructured": "SHURE, M, MOLECULAR-IDENTIFICATION AND ISOLATION OF THE WAXY LOCUS IN MAIZE, CELL 35: 225 (1983).",
      "journal-title": "CELL"
    },
    {
      "key": "e_1_2_1_20_1",
      "doi-asserted-by": "crossref",
      "first-page": "16338",
      "DOI": "10.1016/S0021-9258(17)36242-7",
      "volume": "260",
      "year": "1985",
      "unstructured": "SMART, C.C., SELECTIVE OVERPRODUCTION OF 5-ENOL-PYRUVYLSHIKIMIC ACID 3-PHOSPHATE SYNTHASE IN A PLANT-CELL CULTURE WHICH TOLERATES HIGH-DOSES OF THE HERBICIDE GLYPHOSATE, JOURNAL OF BIOLOGICAL CHEMISTRY 260: 16338 (1985).",
      "journal-title": "JOURNAL OF BIOLOGICAL CHEMISTRY"
    },
    {
      "key": "e_1_2_1_21_1",
      "doi-asserted-by": "crossref",
      "first-page": "169",
      "DOI": "10.1016/0003-9861(86)90106-2",
      "volume": "244",
      "year": "1986",
      "unstructured": "STEINRUCKEN, H.C., OVERPRODUCTION OF 5-ENOLPYRUVYLSHIKIMATE-3-PHOSPHATE SYNTHASE IN A GLYPHOSATE-TOLERANT PETUNIA-HYBRIDA CELL-LINE, ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS 244: 169 (1986).",
      "journal-title": "ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS"
    },
    {
      "key": "e_1_2_1_22_1",
      "doi-asserted-by": "crossref",
      "first-page": "1207",
      "DOI": "10.1016/0006-291X(80)90547-1",
      "volume": "94",
      "year": "1980",
      "unstructured": "STEINRUCKEN, H.C., THE HERBICIDE GLYPHOSATE IS A POTENT INHIBITOR OF 5-ENOLPYRUVYL-SHIKIMIC-ACID 3-PHOSPHATE SYNTHASE, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 94: 1207 (1980).",
      "journal-title": "BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS"
    },
    {
      "key": "e_1_2_1_23_1",
      "first-page": "683",
      "year": "1981",
      "unstructured": "SUGGS, S, ICN UCLA S DEV BIOL: 683 (1981).",
      "journal-title": "ICN UCLA S DEV BIOL"
    },
    {
      "key": "e_1_2_1_24_1",
      "doi-asserted-by": "crossref",
      "first-page": "468",
      "DOI": "10.1016/0076-6879(83)00074-9",
      "volume": "100",
      "year": "1983",
      "unstructured": "ZOLLER, M.J., OLIGONUCLEOTIDE-DIRECTED MUTAGENESIS OF DNA FRAGMENTS CLONED INTO M13 VECTORS, METHODS IN ENZYMOLOGY 100: 468 (1983).",
      "journal-title": "METHODS IN ENZYMOLOGY"
    }
  ],
  "container-title": "Science",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://www.science.org/doi/pdf/10.1126/science.233.4762.478",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2024,
        1,
        12
      ]
    ],
    "date-time": "2024-01-12T16:00:23Z",
    "timestamp": 1705075223000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://www.science.org/doi/10.1126/science.233.4762.478"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        1986,
        7,
        25
      ]
    ]
  },
  "references-count": 24,
  "journal-issue": {
    "issue": "4762",
    "published-print": {
      "date-parts": [
        [
          1986,
          7,
          25
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1126/science.233.4762.478"
  ],
  "URL": "http://dx.doi.org/10.1126/science.233.4762.478",
  "relation": {},
  "ISSN": [
    "0036-8075",
    "1095-9203"
  ],
  "subject": [],
  "container-title-short": "Science",
  "published": {
    "date-parts": [
      [
        1986,
        7,
        25
      ]
    ]
  }
}