DOI: 10.1073/pnas.91.5.1843. Expression and partial purification of enzymaticallyactive recombinant Ty1 integrase in Saccharomyces cerevisiae.

Expression and partial purification of enzymaticallyactive recombinant Ty1 integrase in Saccharomyces cerevisiae.

10.1073/pnas.91.5.1843

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

Integration of the Saccharomyces cerevisiae retrotransposon Ty1 into the genome requires Ty1 integrase (IN). Apparent functions of Ty1 IN are target-site determination, cleavage, and joining of donor strands. To further study the mechanism of Ty1 integration, an IN expression plasmid has been constructed for use in yeast. The recombinant IN coding sequence differs from mature Ty1 IN associated with Ty1 virus-like particles only in that it has several additional N-terminal amino acid codons. Inclusion of a polyhistidine tag facilitates purification of recombinant IN by metal chelate chromatography. Recombinant Ty1 IN is active in an in vitro assay with short double-stranded oligonucleotide substrates and has biochemical properties similar to those observed with Ty1 virus-like particles. The full-length Ty1 IN produced in yeast should be useful for further biochemical, genetic, and structural analyses of Ty1 integration and for comparative analyses with retroviral IN proteins.

Bibliography

Moore, S. P., & Garfinkel, D. J. (1994). Expression and partial purification of enzymaticallyactive recombinant Ty1 integrase in Saccharomyces cerevisiae. Proceedings of the National Academy of Sciences, 91(5), 1843â1847.

Authors 2

S P Moore (first)
D J Garfinkel (additional)

References 0 Referenced 30

None

Dates

Type	When
Created	19 years, 3 months ago (May 31, 2006, 9:05 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 2:04 p.m.)
Indexed	1 year, 7 months ago (Jan. 10, 2024, 5:02 p.m.)
Issued	31 years, 6 months ago (March 1, 1994)
Published	31 years, 6 months ago (March 1, 1994)
Published Online	31 years, 6 months ago (March 1, 1994)
Published Print	31 years, 6 months ago (March 1, 1994)

Funders 0

None

BibTeX

@article{Moore_1994, title={Expression and partial purification of enzymaticallyactive recombinant Ty1 integrase in Saccharomyces cerevisiae.}, volume={91}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.91.5.1843}, DOI={10.1073/pnas.91.5.1843}, number={5}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Moore, S P and Garfinkel, D J}, year={1994}, month=mar, pages={1843–1847} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2024,
        1,
        10
      ]
    ],
    "date-time": "2024-01-10T22:02:32Z",
    "timestamp": 1704924152707
  },
  "reference-count": 0,
  "publisher": "Proceedings of the National Academy of Sciences",
  "issue": "5",
  "content-domain": {
    "domain": [
      "www.pnas.org"
    ],
    "crossmark-restriction": true
  },
  "published-print": {
    "date-parts": [
      [
        1994,
        3
      ]
    ]
  },
  "abstract": "<jats:p>Integration of the Saccharomyces cerevisiae\nretrotransposon Ty1 into the genome requires Ty1 integrase (IN). Apparent\nfunctions of Ty1 IN are target-site determination, cleavage, and joining of\ndonor strands. To further study the mechanism of Ty1 integration, an IN\nexpression plasmid has been constructed for use in yeast. The recombinant IN\ncoding sequence differs from mature Ty1 IN associated with Ty1 virus-like\nparticles only in that it has several additional N-terminal amino acid codons.\nInclusion of a polyhistidine tag facilitates purification of recombinant IN by\nmetal chelate chromatography. Recombinant Ty1 IN is active in an in vitro assay\nwith short double-stranded oligonucleotide substrates and has biochemical\nproperties similar to those observed with Ty1 virus-like particles. The\nfull-length Ty1 IN produced in yeast should be useful for further biochemical,\ngenetic, and structural analyses of Ty1 integration and for comparative analyses\nwith retroviral IN proteins.</jats:p>",
  "DOI": "10.1073/pnas.91.5.1843",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2006,
        5,
        31
      ]
    ],
    "date-time": "2006-05-31T13:05:56Z",
    "timestamp": 1149080756000
  },
  "page": "1843-1847",
  "update-policy": "http://dx.doi.org/10.1073/pnas.cm10313",
  "source": "Crossref",
  "is-referenced-by-count": 30,
  "title": "Expression and partial purification of enzymaticallyactive recombinant Ty1 integrase in Saccharomyces cerevisiae.",
  "prefix": "10.1073",
  "volume": "91",
  "author": [
    {
      "given": "S P",
      "family": "Moore",
      "sequence": "first",
      "affiliation": [
        {
          "name": "National Cancer Institute-Frederick Cancer Research and\rDevelopment Center Advanced BioScience Laboratories-Basic Research Program,\rFrederick, MD 21702-1201."
        }
      ]
    },
    {
      "given": "D J",
      "family": "Garfinkel",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "National Cancer Institute-Frederick Cancer Research and\rDevelopment Center Advanced BioScience Laboratories-Basic Research Program,\rFrederick, MD 21702-1201."
        }
      ]
    }
  ],
  "member": "341",
  "published-online": {
    "date-parts": [
      [
        1994,
        3
      ]
    ]
  },
  "container-title": "Proceedings of the National Academy of Sciences",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://pnas.org/doi/pdf/10.1073/pnas.91.5.1843",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2022,
        4,
        13
      ]
    ],
    "date-time": "2022-04-13T18:04:50Z",
    "timestamp": 1649873090000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://pnas.org/doi/full/10.1073/pnas.91.5.1843"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        1994,
        3
      ]
    ]
  },
  "references-count": 0,
  "journal-issue": {
    "issue": "5",
    "published-print": {
      "date-parts": [
        [
          1994,
          3
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1073/pnas.91.5.1843"
  ],
  "URL": "http://dx.doi.org/10.1073/pnas.91.5.1843",
  "relation": {},
  "ISSN": [
    "0027-8424",
    "1091-6490"
  ],
  "subject": [],
  "container-title-short": "Proc. Natl. Acad. Sci. U.S.A.",
  "published": {
    "date-parts": [
      [
        1994,
        3
      ]
    ]
  },
  "assertion": [
    {
      "value": "1994-03-01",
      "order": 2,
      "name": "published",
      "label": "Published",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    }
  ]
}