Expression of human adenosine deaminase using a transmissable murine retrovirus vector system
- PMID: 3856223
- PMCID: PMC397114
- DOI: 10.1073/pnas.82.3.703
Expression of human adenosine deaminase using a transmissable murine retrovirus vector system
Abstract
Human adenosine deaminase (ADA; adenosine aminohydrolase, EC 3.5.4.4) was expressed at high levels in cultured mouse cells using a transmissable murine retrovirus vector system. A cDNA clone encoding ADA has been inserted into a plasmid vector containing retroviral transcription and packaging signals as well as a selectable gene for G418 resistance. The constructions were transfected into psi 2 cells, which package the recombinant retroviral genomes into replication-defective virus particles. Isoenzyme analysis for ADA in G418-selected psi 2 cells showed at least 20-fold more human ADA activity than endogenous mouse ADA activity. A mouse T-cell lymphoma line, BL/VL3, was cocultured with transformed psi 2 cells producing human ADA, and some of the cocultured cells were selected for resistance to G418. Both G418-selected and unselected cocultured cells expressed human ADA activity at 25%-50% the level of the endogenous enzyme. Thus, efficient retroviral transduction of ADA expression was obtained.
Similar articles
-
Retrovirus-mediated transfer of human adenosine deaminase gene sequences into cells in culture and into murine hematopoietic cells in vivo.Proc Natl Acad Sci U S A. 1986 Apr;83(8):2566-70. doi: 10.1073/pnas.83.8.2566. Proc Natl Acad Sci U S A. 1986. PMID: 3458218 Free PMC article.
-
Introduction of sequences encoding functional human adenosine deaminase into mouse cells using a retroviral shuttle system.Gene. 1985;34(2-3):163-8. doi: 10.1016/0378-1119(85)90124-6. Gene. 1985. PMID: 4007492
-
Efficient retrovirus-mediated transfer and expression of a human adenosine deaminase gene in diploid skin fibroblasts from an adenosine deaminase-deficient human.Proc Natl Acad Sci U S A. 1987 Feb;84(4):1055-9. doi: 10.1073/pnas.84.4.1055. Proc Natl Acad Sci U S A. 1987. PMID: 3493485 Free PMC article.
-
Retroviral-mediated gene transfer into mammalian cells.Blood Cells. 1987;13(1-2):285-98. Blood Cells. 1987. PMID: 3311223 Review.
-
Bone marrow gene therapy for adenosine deaminase deficiency.Immunodeficiency. 1993;4(1-4):335-45. Immunodeficiency. 1993. PMID: 7909479 Review.
Cited by
-
Retrovirus-mediated transfer of human adenosine deaminase gene sequences into cells in culture and into murine hematopoietic cells in vivo.Proc Natl Acad Sci U S A. 1986 Apr;83(8):2566-70. doi: 10.1073/pnas.83.8.2566. Proc Natl Acad Sci U S A. 1986. PMID: 3458218 Free PMC article.
-
Correction of adenosine deaminase deficiency in cultured human T and B cells by retrovirus-mediated gene transfer.Proc Natl Acad Sci U S A. 1986 Sep;83(17):6563-7. doi: 10.1073/pnas.83.17.6563. Proc Natl Acad Sci U S A. 1986. PMID: 3489233 Free PMC article.
-
In vitro development of B lymphocytes from long-term cultured precursor cells.Proc Natl Acad Sci U S A. 1986 Jan;83(2):441-5. doi: 10.1073/pnas.83.2.441. Proc Natl Acad Sci U S A. 1986. PMID: 3079911 Free PMC article.
-
High mutation rate of a spleen necrosis virus-based retrovirus vector.Mol Cell Biol. 1986 Dec;6(12):4387-95. doi: 10.1128/mcb.6.12.4387-4395.1986. Mol Cell Biol. 1986. PMID: 3796606 Free PMC article.
-
Retroviral-mediated gene transfer. Applications in neurobiology.Mol Neurobiol. 1988 Fall;2(3):155-82. doi: 10.1007/BF02935344. Mol Neurobiol. 1988. PMID: 2855793 Review.
References
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Full Text Sources
Other Literature Sources
Research Materials
