Human adenovirus cloning vectors based on infectious bacterial plasmids

Goutam Ghosh-Choudhury; Yousef Haj-Ahmad; Pamela Brinkley; John Rudy; Frank L. Graham

doi:10.1016/0378-1119(86)90321-5

Human adenovirus cloning vectors based on infectious bacterial plasmids

Goutam Ghosh-Choudhury, Yousef Haj-Ahmad, Pamela Brinkley, John Rudy, Frank L. Graham

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

59 Scopus citations

Abstract

By making use of the fact that human adenovirus DNA circularizes in infected cells, and that circular forms of the viral genome are infectious, we have developed an improved adenovirus-based cloning system. A deletion mutant of adenovirus type 5 (Ad5) with deletions in early regions 1 (E1) and 3 (E3) was converted to a bacterial plasmid which can regenerate infectious virus following transfection into human 293 cells. A single XbaI recognition site in the deleted E3 region serves as a site for the insertion of foreign DNA. We have used this system to clone a number of genes into the Ad5 genome and describe the insertion of the neomycin/G418 resistance marker into Ad5 as an example.

Original language	English (US)
Pages (from-to)	161-171
Number of pages	11
Journal	Gene
Volume	50
Issue number	1-3
DOIs	https://doi.org/10.1016/0378-1119(86)90321-5
State	Published - 1986
Externally published	Yes

Keywords

Ad5 insertion mutants
DNA transfection
Recombinant DNA
bacteriophage λ cos site
gene transfer
neomycin resistance
shuttle vectors

ASJC Scopus subject areas

Genetics

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10.1016/0378-1119(86)90321-5

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title = "Human adenovirus cloning vectors based on infectious bacterial plasmids",

abstract = "By making use of the fact that human adenovirus DNA circularizes in infected cells, and that circular forms of the viral genome are infectious, we have developed an improved adenovirus-based cloning system. A deletion mutant of adenovirus type 5 (Ad5) with deletions in early regions 1 (E1) and 3 (E3) was converted to a bacterial plasmid which can regenerate infectious virus following transfection into human 293 cells. A single XbaI recognition site in the deleted E3 region serves as a site for the insertion of foreign DNA. We have used this system to clone a number of genes into the Ad5 genome and describe the insertion of the neomycin/G418 resistance marker into Ad5 as an example.",

keywords = "Ad5 insertion mutants, DNA transfection, Recombinant DNA, bacteriophage λ cos site, gene transfer, neomycin resistance, shuttle vectors",

author = "Goutam Ghosh-Choudhury and Yousef Haj-Ahmad and Pamela Brinkley and John Rudy and Graham, {Frank L.}",

note = "Funding Information: We wish to thank Silvia Bacchettif or helpful commentsa nd Janice Butera and Barb Lahie for typingt hem anuscriptT. his work was supportedb y grantsfr omt heN ationalC ancerI nstituteo f Canada and the Medical Research Council of Canada. F.L.G. is a Terry Fox CancerR esearch Scientoisf t the National CancerI nstituteo f Canada.",

year = "1986",

doi = "10.1016/0378-1119(86)90321-5",

language = "English (US)",

volume = "50",

pages = "161--171",

journal = "Gene",

issn = "0378-1119",

publisher = "Elsevier",

number = "1-3",

}

TY - JOUR

T1 - Human adenovirus cloning vectors based on infectious bacterial plasmids

AU - Ghosh-Choudhury, Goutam

AU - Haj-Ahmad, Yousef

AU - Brinkley, Pamela

AU - Rudy, John

AU - Graham, Frank L.

N1 - Funding Information: We wish to thank Silvia Bacchettif or helpful commentsa nd Janice Butera and Barb Lahie for typingt hem anuscriptT. his work was supportedb y grantsfr omt heN ationalC ancerI nstituteo f Canada and the Medical Research Council of Canada. F.L.G. is a Terry Fox CancerR esearch Scientoisf t the National CancerI nstituteo f Canada.

PY - 1986

Y1 - 1986

N2 - By making use of the fact that human adenovirus DNA circularizes in infected cells, and that circular forms of the viral genome are infectious, we have developed an improved adenovirus-based cloning system. A deletion mutant of adenovirus type 5 (Ad5) with deletions in early regions 1 (E1) and 3 (E3) was converted to a bacterial plasmid which can regenerate infectious virus following transfection into human 293 cells. A single XbaI recognition site in the deleted E3 region serves as a site for the insertion of foreign DNA. We have used this system to clone a number of genes into the Ad5 genome and describe the insertion of the neomycin/G418 resistance marker into Ad5 as an example.

AB - By making use of the fact that human adenovirus DNA circularizes in infected cells, and that circular forms of the viral genome are infectious, we have developed an improved adenovirus-based cloning system. A deletion mutant of adenovirus type 5 (Ad5) with deletions in early regions 1 (E1) and 3 (E3) was converted to a bacterial plasmid which can regenerate infectious virus following transfection into human 293 cells. A single XbaI recognition site in the deleted E3 region serves as a site for the insertion of foreign DNA. We have used this system to clone a number of genes into the Ad5 genome and describe the insertion of the neomycin/G418 resistance marker into Ad5 as an example.

KW - Ad5 insertion mutants

KW - DNA transfection

KW - Recombinant DNA

KW - bacteriophage λ cos site

KW - gene transfer

KW - neomycin resistance

KW - shuttle vectors

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U2 - 10.1016/0378-1119(86)90321-5

DO - 10.1016/0378-1119(86)90321-5

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AN - SCOPUS:0022884352

SN - 0378-1119

VL - 50

SP - 161

EP - 171

JO - Gene

JF - Gene

IS - 1-3

ER -

Human adenovirus cloning vectors based on infectious bacterial plasmids

Abstract

Keywords

ASJC Scopus subject areas

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