Literature DB >> 6170119

Efficient DNA transfection and rapid assay for thymidine kinase activity and viral antigenic determinants.

G Milman, M Herzberg.   

Abstract

We describe DEAE dextran-mediated DNA transfection in suspension which routinely gives transient gene expression in 0.1--1% of the transfected cells. We have used normal diploid human skin fibroblasts, monkey BSC cells, and mouse L or 3T6 cells with almost equal efficiency. Gene expression is detected 1--3 days after addition of the DNA. SV40 and polyoma T and V antigen are detected by in situ immunofluorescence and thymidine kinase gene expression is detected by in situ autoradiography. The high efficiency of transfection and the speed of detection together provide a means to study transfecting gene functions that does not rely on selection to obtain stable transformants. It should be possible to screen for the expression of any gene product which can be assayed by in situ immunofluorescence or autoradiographic techniques.

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Year:  1981        PMID: 6170119     DOI: 10.1007/BF01567655

Source DB:  PubMed          Journal:  Somatic Cell Genet        ISSN: 0098-0366


  22 in total

1.  Acquired cisplatin resistance in human ovarian cancer cells is associated with enhanced repair of cisplatin-DNA lesions and reduced drug accumulation.

Authors:  R J Parker; A Eastman; F Bostick-Bruton; E Reed
Journal:  J Clin Invest       Date:  1991-03       Impact factor: 14.808

2.  High-frequency transfer of cloned herpes simplex virus type 1 sequences to mammalian cells by protoplast fusion.

Authors:  R M Sandri-Goldin; A L Goldin; M Levine; J C Glorioso
Journal:  Mol Cell Biol       Date:  1981-08       Impact factor: 4.272

3.  Linear simian virus 40 DNA fragments exhibit a propensity for rolling-circle replication.

Authors:  I Deichaite; Z Laver-Rudich; D Dorsett; E Winocour
Journal:  Mol Cell Biol       Date:  1985-07       Impact factor: 4.272

4.  Expression of transfected DNA in avian cells can be enhanced in trans by retroviral infection.

Authors:  B R Cullen; R A Katz; G Ju
Journal:  Mol Cell Biol       Date:  1985-07       Impact factor: 4.272

5.  Simian-human immunodeficiency virus (SHIV) containing the nef/long terminal repeat region of the highly virulent SIVsmmPBj14 causes PBj-like activation of cultured resting peripheral blood mononuclear cells, but the chimera showed No increase in virulence.

Authors:  E B Stephens; S Mukherjee; Z Q Liu; D Sheffer; R Lamb-Wharton; K Leung; W Zhuge; S V Joag; Z Li; L Foresman; I Adany; O Narayan
Journal:  J Virol       Date:  1998-06       Impact factor: 5.103

6.  Short-term, high-efficiency expression of transfected DNA.

Authors:  D J Sussman; G Milman
Journal:  Mol Cell Biol       Date:  1984-08       Impact factor: 4.272

7.  Efficient transfer of cloned DNA into human diploid cells: protoplast fusion in suspension.

Authors:  P Litzkas; K K Jha; H L Ozer
Journal:  Mol Cell Biol       Date:  1984-11       Impact factor: 4.272

8.  Comparison of phenotypic expression with genotypic transformation by using cloned, selectable markers.

Authors:  P S Linsley; L Siminovitch
Journal:  Mol Cell Biol       Date:  1982-05       Impact factor: 4.272

9.  Identification of human genomic clones coding the major histocompatibility antigens HLA-a2 and HLA-B7 by DNA-mediated gene transfer.

Authors:  J A Barbosa; M E Kamarck; P A Biro; S M Weissman; F H Ruddle
Journal:  Proc Natl Acad Sci U S A       Date:  1982-10       Impact factor: 11.205

10.  Expression of cloned herpesvirus genes. I. Detection of nuclear antigens from herpes simplex virus type 2 inverted repeat regions in transfected mouse cells.

Authors:  M H Middleton; G R Reyes; D M Ciufo; A Buchan; J C Macnab; G S Hayward
Journal:  J Virol       Date:  1982-09       Impact factor: 5.103

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