Home LiteratureArticle Details

PMID: 1720540 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

A dominant positive and negative selectable gene for use in mammalian cells.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 88 ·No. 23 ·1991-12-01 ·Pages 10416-20

Schwartz F, Maeda N, Smithies O, Hickey R, Edelmann W, Skoultchi A, Kucherlapati R

Abstract

We have constructed three different fusion genes containing the herpes simplex virus thymidine kinase (HSV tk) and the bacterial neomycin phosphotransferase (neo) genes. All three fusion genes utilize the HSV tk promoter but differ at the junction of their components. We have determined if the fusion genes are bifunctional by introducing them into mammalian cells and testing for function of the individual components. One of the fusion genes, TNFUS 69, produced a bicistronic message and a fusion protein that has TK and NEO protein functions. This and other fusion genes of a similar nature could serve as dominant positive and negative selectable markers in mammalian cells.

Related Genes

neo

MeSH Terms

Amino Acid Sequence Animals Bacteria/enzymology,genetics Base Sequence Blotting, Northern Cloning, Molecular/methods Genes, Bacterial Genes, Dominant Genetic Markers Kanamycin Kinase L Cells Mice Molecular Sequence Data Phosphotransferases/genetics Plasmids Promoter Regions, Genetic RNA/genetics,isolation & purification Restriction Mapping Simplexvirus/enzymology,genetics Thymidine Kinase/genetics Transfection

Chemicals

Genetic Markers RNA Phosphotransferases Thymidine Kinase Kanamycin Kinase

Authors & Affiliations

7 authors, click to expand affiliations / ORCID

Schwartz F

Department of Genetics, University of Illinois College of Medicine, Chicago 60612.

Maeda N

Smithies O

Hickey R

Edelmann W

Skoultchi A

Kucherlapati R

References (21)

21 references, click to expand

DELETION OF THYMIDINE KINASE ACTIVITY FROM L CELLS RESISTANT TO BROMODEOXYURIDINE.
Exp Cell Res. 1963 Aug;31:297-312 PMID: 14056989
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
Dominant positive and negative selection using a hygromycin phosphotransferase-thymidine kinase fusion gene.
Mol Cell Biol. 1991 Jun;11(6):3374-8 PMID: 1645450
Testing an "in-out" targeting procedure for making subtle genomic modifications in mouse embryonic stem cells.
Mol Cell Biol. 1991 Mar;11(3):1402-8 PMID: 1996101
Positive genetic selection for gene disruption in mammalian cells by homologous recombination.
Proc Natl Acad Sci U S A. 1989 Jan;86(1):227-31 PMID: 2536156
Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells.
Cell. 1987 Nov 6;51(3):503-12 PMID: 2822260
Disruption of the proto-oncogene int-2 in mouse embryo-derived stem cells: a general strategy for targeting mutations to non-selectable genes.
Nature. 1988 Nov 24;336(6197):348-52 PMID: 3194019
Protein fusions with the kanamycin resistance gene from transposon Tn5.
EMBO J. 1984 Dec 20;3(13):3317-22 PMID: 6098471
Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose.
Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201-5 PMID: 6159641
Analysis of transcriptional regulatory signals of the HSV thymidine kinase gene: identification of an upstream control region.
Cell. 1981 Aug;25(2):385-98 PMID: 6269744
Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter.
J Mol Appl Genet. 1982;1(4):327-41 PMID: 6286831
The nucleotide sequence and transcript map of the herpes simplex virus thymidine kinase gene.
Nucleic Acids Res. 1980 Dec 20;8(24):5949-64 PMID: 6258156
Expression of a bacterial gene in mammalian cells.
Science. 1980 Sep 19;209(4463):1422-7 PMID: 6251549
Transfer of purified herpes virus thymidine kinase gene to cultured mouse cells.
Cell. 1977 May;11(1):223-32 PMID: 194704
Helper-independent transformation by unintegrated Harvey sarcoma virus DNA.
J Virol. 1978 May;26(2):291-8 PMID: 26810
Cloning of the active thymidine kinase gene of herpes simplex virus type 1 in Escherichia coli K-12.
Proc Natl Acad Sci U S A. 1979 Aug;76(8):3755-9 PMID: 226978
Biochemical transfer of single-copy eucaryotic genes using total cellular DNA as donor.
Cell. 1978 Jul;14(3):725-31 PMID: 210957
Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4 PMID: 388439
Plasmid-determined resistance to antimicrobial agents.
Annu Rev Microbiol. 1978;32:469-518 PMID: 360974
Competition hybridization by "pre-saturation" of HeLa cell DNA.
J Mol Biol. 1969 Sep 28;44(3):551-62 PMID: 5388124
Introduction of a subtle mutation into the Hox-2.6 locus in embryonic stem cells.
Nature. 1991 Mar 21;350(6315):243-6 PMID: 1672446

Article Info

Journal

Proceedings of the National Academy of Sciences of the United States of America

Abbr.

Proc Natl Acad Sci U S A

ISSN

0027-8424

DOI

10.1073/pnas.88.23.10416

Published

1991-12-01

Pages

10416-20

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC52939

Subset

Grants

NIGMS NIH HHS · GM20069 · United States

NHLBI NIH HHS · HL37001 · United States

NHLBI NIH HHS · HL42630 · United States

External Links

PubMed source DOI: 10.1073/pnas.88.23.10416 PMC Full Text View journal details

Analysis Services

A dominant positive and negative selectable gene for use in mammalian cells.

Contact

WeChat Official Account