One-step gene amplification by Mu-mediated transposition of E. coli genes to a multicopy plasmid

Diego de Mendoza; David Clark; John E. Cronan

doi:10.1016/0378-1119(81)90101-3

One-step gene amplification by Mu-mediated transposition of E. coli genes to a multicopy plasmid

Diego de Mendoza, David Clark, John E. Cronan

Research output: Contribution to journal › Article › peer-review

Abstract

A general in vivo method to amplify the number of copies of a specific gene in one step is described. The method is directly applicable to any selectable gene of Escherichia coli and is based on the Mu-mediated transposition of segments of host chromosomes into the conjugative, multicopy plasmid R6K. Using this method we have cloned the β-hydroxydecanoyl thioester dehydrase structural gene, fabA, into the R6K plasmid. Strains carrying the resultant plasmid produced 13 to 21 times more dehydrase than control strains.

Original language	English (US)
Pages (from-to)	27-32
Number of pages	6
Journal	Gene
Volume	15
Issue number	1
DOIs	https://doi.org/10.1016/0378-1119(81)90101-3
State	Published - Oct 1981

Keywords

Molecular cloning
R6K vector
fabA
in vivo genetic engineering
β-hydroxydecanoyl thioester dehydrase

ASJC Scopus subject areas

Genetics

Online availability

10.1016/0378-1119(81)90101-3

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title = "One-step gene amplification by Mu-mediated transposition of E. coli genes to a multicopy plasmid",

abstract = "A general in vivo method to amplify the number of copies of a specific gene in one step is described. The method is directly applicable to any selectable gene of Escherichia coli and is based on the Mu-mediated transposition of segments of host chromosomes into the conjugative, multicopy plasmid R6K. Using this method we have cloned the β-hydroxydecanoyl thioester dehydrase structural gene, fabA, into the R6K plasmid. Strains carrying the resultant plasmid produced 13 to 21 times more dehydrase than control strains.",

keywords = "Molecular cloning, R6K vector, fabA, in vivo genetic engineering, β-hydroxydecanoyl thioester dehydrase",

author = "{de Mendoza}, Diego and David Clark and Cronan, {John E.}",

note = "Funding Information: We thank Dr. M. Howe for gifts of phagea ndbac-terial strains.T his work was supportedb y NIH grants GM 26156 ar,d NSF grant 79-25689a nd a postdoctoral fellowship( to DdM) from the ConsejoN acional de InvestigacionesC ientificas y T6cmcas de la RepdblicaA rgentina.",

year = "1981",

month = oct,

doi = "10.1016/0378-1119(81)90101-3",

language = "English (US)",

volume = "15",

pages = "27--32",

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T1 - One-step gene amplification by Mu-mediated transposition of E. coli genes to a multicopy plasmid

AU - de Mendoza, Diego

AU - Clark, David

AU - Cronan, John E.

N1 - Funding Information: We thank Dr. M. Howe for gifts of phagea ndbac-terial strains.T his work was supportedb y NIH grants GM 26156 ar,d NSF grant 79-25689a nd a postdoctoral fellowship( to DdM) from the ConsejoN acional de InvestigacionesC ientificas y T6cmcas de la RepdblicaA rgentina.

PY - 1981/10

Y1 - 1981/10

N2 - A general in vivo method to amplify the number of copies of a specific gene in one step is described. The method is directly applicable to any selectable gene of Escherichia coli and is based on the Mu-mediated transposition of segments of host chromosomes into the conjugative, multicopy plasmid R6K. Using this method we have cloned the β-hydroxydecanoyl thioester dehydrase structural gene, fabA, into the R6K plasmid. Strains carrying the resultant plasmid produced 13 to 21 times more dehydrase than control strains.

AB - A general in vivo method to amplify the number of copies of a specific gene in one step is described. The method is directly applicable to any selectable gene of Escherichia coli and is based on the Mu-mediated transposition of segments of host chromosomes into the conjugative, multicopy plasmid R6K. Using this method we have cloned the β-hydroxydecanoyl thioester dehydrase structural gene, fabA, into the R6K plasmid. Strains carrying the resultant plasmid produced 13 to 21 times more dehydrase than control strains.

KW - Molecular cloning

KW - R6K vector

KW - fabA

KW - in vivo genetic engineering

KW - β-hydroxydecanoyl thioester dehydrase

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JO - Gene

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One-step gene amplification by Mu-mediated transposition of E. coli genes to a multicopy plasmid

Abstract

Keywords

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