Home LiteratureArticle Details
PMID: 6417663 Published · ppublish English Journal Article

Transient expression of genes introduced into cultured cells of Drosophila.

Di Nocera PP, Dawid IB

Abstract

Recombinant plasmids in which the sequence encoding the bacterial chloramphenicol acetyltransferase (CAT; acetyl-CoA:chloramphenicol 3-O-acetyltransferase, EC 2.3.1.28) has been placed under the control of Drosophila heat shock protein 70 (hsp 70) or copia promoters have been introduced into cultured cells of two Drosophila species (Schneider II line of Drosophila melanogaster and D. immigrans) as calcium-phosphate complexes. Within 1-2 days after transfection functional CAT enzyme was detected in cells exposed to either CAT recombinant. The expression of the bacterial information depends on the activity of the Drosophila promoters because plasmids in which the Drosophila DNA fragments were fused to the CAT coding sequence in inverted orientation did not support the synthesis of CAT enzyme activity. Low levels of CAT activity and of hybrid mRNA were detected in cells transformed with hsp-cat recombinants when the cells were maintained at room temperature, and both mRNA levels and CAT activity increased substantially after a brief exposure to 37 degrees C. hsp-cat mRNA has the same 5' terminus as authentic Drosophila hsp 70 messenger. These experiments document a practical system for the introduction and expression of isolated genes in cultured cells of Drosophila.

MeSH Terms
Acetyltransferases/genetics Base Sequence Cells, Cultured Chloramphenicol O-Acetyltransferase DNA, Recombinant/metabolism Drosophila/enzymology,genetics Drosophila melanogaster/enzymology,genetics Genes Plasmids Transfection
Chemicals
DNA, Recombinant Acetyltransferases Chloramphenicol O-Acetyltransferase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Di Nocera P P
Dawid I B
References (27)
27 references, click to expand
  1. Direct transfer of cloned genes from bacteria to mammalian cells.
    Proc Natl Acad Sci U S A. 1980 Apr;77(4):2163-7 PMID: 6246525
  2. Four heat shock proteins of Drosophila melanogaster coded within a 12-kilobase region in chromosome subdivision 67B.
    Proc Natl Acad Sci U S A. 1980 Sep;77(9):5390-3 PMID: 6254079
  3. The 5' termini of RNAs encoded by the transposable element copia.
    Nucleic Acids Res. 1981 Dec 11;9(23):6279-91 PMID: 6275356
  4. Vectors containing a prokaryotic dihydrofolate reductase gene transform Drosophila cells to methotrexate-resistance.
    EMBO J. 1983;2(7):1099-104 PMID: 6194991
  5. Nucleotide sequences of heat shock activated genes in Drosophila melanogaster. I. Sequences in the regions of the 5' and 3' ends of the hsp 70 gene in the hybrid plasmid 56H8.
    Nucleic Acids Res. 1980 Jul 25;8(14):3105-23 PMID: 6255408
  6. Regulated transcription of the genes for actin and heat-shock proteins in cultured Drosophila cells.
    J Cell Biol. 1981 Feb;88(2):323-8 PMID: 6162853
  7. High efficiency transformation by direct microinjection of DNA into cultured mammalian cells.
    Cell. 1980 Nov;22(2 Pt 2):479-88 PMID: 6256082
  8. Heat-shock proteins are associated with hnRNA in Drosophila melanogaster tissue culture cells.
    EMBO J. 1983;2(5):705-10 PMID: 16453451
  9. The Rous sarcoma virus long terminal repeat is a strong promoter when introduced into a variety of eukaryotic cells by DNA-mediated transfection.
    Proc Natl Acad Sci U S A. 1982 Nov;79(22):6777-81 PMID: 6294651
  10. Cell lines derived from late embryonic stages of Drosophila melanogaster.
    J Embryol Exp Morphol. 1972 Apr;27(2):353-65 PMID: 4625067
  11. Gene transfer in amphibian eggs and oocytes.
    Annu Rev Genet. 1981;15:189-218 PMID: 7039494
  12. Translational efficiency of heat-induced messages in Drosophila melanogaster cells.
    J Mol Biol. 1980 Feb 25;137(2):151-8 PMID: 6768893
  13. Translational control of protein synthesis in response to heat shock in D. melanogaster cells.
    Cell. 1980 Dec;22(3):825-34 PMID: 6780199
  14. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells.
    Mol Cell Biol. 1982 Sep;2(9):1044-51 PMID: 6960240
  15. Rat insulin genes: construction of plasmids containing the coding sequences.
    Science. 1977 Jun 17;196(4296):1313-9 PMID: 325648
  16. Transformation of yeast.
    Proc Natl Acad Sci U S A. 1978 Apr;75(4):1929-33 PMID: 347451
  17. A rapid alkaline extraction procedure for screening recombinant plasmid DNA.
    Nucleic Acids Res. 1979 Nov 24;7(6):1513-23 PMID: 388356
  18. Stimulation by cyclic adenosine monophosphate of plasmid deoxyribonucleic acid replication and catabolite repression of the plasmid deoxyribonucleic acid-protein relaxation complex.
    J Bacteriol. 1973 May;114(2):577-91 PMID: 4350343
  19. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene.
    Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961-5 PMID: 1105573
  20. Heat shock and recovery are mediated by different translational mechanisms.
    Proc Natl Acad Sci U S A. 1982 Oct;79(20):6181-5 PMID: 6815647
  21. Repeated gene families in Drosophila melanogaster.
    Cold Spring Harb Symp Quant Biol. 1978;42 Pt 2:1053-63 PMID: 98263
  22. Calcium-dependent bacteriophage DNA infection.
    J Mol Biol. 1970 Oct 14;53(1):159-62 PMID: 4922220
  23. Autonomous replication and expression of RNA 1 from black beetle virus.
    J Virol. 1983 May;46(2):481-9 PMID: 16789241
  24. Studies on the selectivity of DNA precipitation by spermine.
    Nucleic Acids Res. 1981 Oct 24;9(20):5493-504 PMID: 7029471
  25. Sequencing end-labeled DNA with base-specific chemical cleavages.
    Methods Enzymol. 1980;65(1):499-560 PMID: 6246368
  26. A new technique for the assay of infectivity of human adenovirus 5 DNA.
    Virology. 1973 Apr;52(2):456-67 PMID: 4705382
  27. A regulatory upstream promoter element in the Drosophila hsp 70 heat-shock gene.
    Cell. 1982 Sep;30(2):517-28 PMID: 6814763
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
Published
1983-12-00
Pages
7095-8
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC389999
Subset
IM
Databases
GENBANK
K01720
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com