A plasmid vector with positive selection and directional cloning based on a conditionally lethal gene

Sergey A. Yazynin, Sergey M. Deyev, Milan Jucovič, Robert W. Hartley

    Research output: Contribution to journalArticle

    22 Citations (Scopus)

    Abstract

    A plasmid vector with a multiple cloning site (MCS) for positive selection of cloned inserts in Escherichia coli (Ec) has been devised, based on the expression plasmid (pMT416) for the bacterial ribonuclease barnase (Barn). The host is protected from the lethal effect of moderate expression of barn by expression of the gene bars, encoding its inhibitor, barstar (Bars), placed on the same plasmid. Full expression, however, is lethal. Induction is also lethal with the derived plasmid, pMT440, which has the pUC19 MCS inserted into barn. Under inducing conditions, transformation by the vector is lethal unless the product of the modified barn is inactivated by insertion of cloned DNA fragments into the MCS. Plasmid pMT440 is, therefore, a generally useful selective cloning vector not requiring any special strain of Ec.

    Original languageEnglish
    Pages (from-to)131-132
    Number of pages2
    JournalGene
    Volume169
    Issue number1
    DOIs
    Publication statusPublished - 22 Feb 1996

    Fingerprint

    Lethal Genes
    Organism Cloning
    Plasmids
    Escherichia coli
    Genetic Vectors
    Ribonucleases
    Gene Expression
    DNA

    Keywords

    • Barnase
    • Barstar
    • Polylinker
    • Ribonuclease

    ASJC Scopus subject areas

    • Medicine(all)
    • Genetics

    Cite this

    A plasmid vector with positive selection and directional cloning based on a conditionally lethal gene. / Yazynin, Sergey A.; Deyev, Sergey M.; Jucovič, Milan; Hartley, Robert W.

    In: Gene, Vol. 169, No. 1, 22.02.1996, p. 131-132.

    Research output: Contribution to journalArticle

    Yazynin, Sergey A. ; Deyev, Sergey M. ; Jucovič, Milan ; Hartley, Robert W. / A plasmid vector with positive selection and directional cloning based on a conditionally lethal gene. In: Gene. 1996 ; Vol. 169, No. 1. pp. 131-132.
    @article{ad20bf9c6b02403b924b866bce2c2bbf,
    title = "A plasmid vector with positive selection and directional cloning based on a conditionally lethal gene",
    abstract = "A plasmid vector with a multiple cloning site (MCS) for positive selection of cloned inserts in Escherichia coli (Ec) has been devised, based on the expression plasmid (pMT416) for the bacterial ribonuclease barnase (Barn). The host is protected from the lethal effect of moderate expression of barn by expression of the gene bars, encoding its inhibitor, barstar (Bars), placed on the same plasmid. Full expression, however, is lethal. Induction is also lethal with the derived plasmid, pMT440, which has the pUC19 MCS inserted into barn. Under inducing conditions, transformation by the vector is lethal unless the product of the modified barn is inactivated by insertion of cloned DNA fragments into the MCS. Plasmid pMT440 is, therefore, a generally useful selective cloning vector not requiring any special strain of Ec.",
    keywords = "Barnase, Barstar, Polylinker, Ribonuclease",
    author = "Yazynin, {Sergey A.} and Deyev, {Sergey M.} and Milan Jucovič and Hartley, {Robert W.}",
    year = "1996",
    month = "2",
    day = "22",
    doi = "10.1016/0378-1119(95)00814-4",
    language = "English",
    volume = "169",
    pages = "131--132",
    journal = "Gene",
    issn = "0378-1119",
    publisher = "Elsevier",
    number = "1",

    }

    TY - JOUR

    T1 - A plasmid vector with positive selection and directional cloning based on a conditionally lethal gene

    AU - Yazynin, Sergey A.

    AU - Deyev, Sergey M.

    AU - Jucovič, Milan

    AU - Hartley, Robert W.

    PY - 1996/2/22

    Y1 - 1996/2/22

    N2 - A plasmid vector with a multiple cloning site (MCS) for positive selection of cloned inserts in Escherichia coli (Ec) has been devised, based on the expression plasmid (pMT416) for the bacterial ribonuclease barnase (Barn). The host is protected from the lethal effect of moderate expression of barn by expression of the gene bars, encoding its inhibitor, barstar (Bars), placed on the same plasmid. Full expression, however, is lethal. Induction is also lethal with the derived plasmid, pMT440, which has the pUC19 MCS inserted into barn. Under inducing conditions, transformation by the vector is lethal unless the product of the modified barn is inactivated by insertion of cloned DNA fragments into the MCS. Plasmid pMT440 is, therefore, a generally useful selective cloning vector not requiring any special strain of Ec.

    AB - A plasmid vector with a multiple cloning site (MCS) for positive selection of cloned inserts in Escherichia coli (Ec) has been devised, based on the expression plasmid (pMT416) for the bacterial ribonuclease barnase (Barn). The host is protected from the lethal effect of moderate expression of barn by expression of the gene bars, encoding its inhibitor, barstar (Bars), placed on the same plasmid. Full expression, however, is lethal. Induction is also lethal with the derived plasmid, pMT440, which has the pUC19 MCS inserted into barn. Under inducing conditions, transformation by the vector is lethal unless the product of the modified barn is inactivated by insertion of cloned DNA fragments into the MCS. Plasmid pMT440 is, therefore, a generally useful selective cloning vector not requiring any special strain of Ec.

    KW - Barnase

    KW - Barstar

    KW - Polylinker

    KW - Ribonuclease

    UR - http://www.scopus.com/inward/record.url?scp=0029931855&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=0029931855&partnerID=8YFLogxK

    U2 - 10.1016/0378-1119(95)00814-4

    DO - 10.1016/0378-1119(95)00814-4

    M3 - Article

    VL - 169

    SP - 131

    EP - 132

    JO - Gene

    JF - Gene

    SN - 0378-1119

    IS - 1

    ER -