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A Human Parvovirus, Adeno-Associated Virus, as a Eucaryotic Vector: Transient Expression and Encapsidation of the Procaryotic Gene for Chloramphenicol Acetyltransferase
Open accessAbstract
We have used the defective human parvovirus adeno-associated virus (AAV) as a novel eucaryotic vector (parvector) for the expression of a foreign gene in human cells. The recombinant, pAV2, contains the AAV genome in a pBR322-derived bacterial plasmid. When pAV2 is transfected into human cells together with helper adenovirus particles, the AAV genome is rescued from the recombinant plasmid and replicated to produce infectious AAV particles at high efficiency. To create a vector, we inserted a procaryotic sequence coding for chloramphenicol acetyltransferase (CAT) into derivatives of pAV2 following either of the AAV promoters p40 (pAVHiCAT) and p19 (pAVBcCAT). When transfected into human 293 cells or HeLa cells, pAVHiCAT expressed CAT activity in the absence of adenovirus. In the presence of adenovirus, this vector produced increased amounts of CAT activity and the recombinant AAV-CAT genome was replicated. In 293 cells, pAVBcCAT expressed a similar amount of CAT activity in the absence or presence of adenovirus and the recombinant AAV-CAT genome was not replicated. In HeLa cells, pAVBcCAT expressed low levels of CAT activity, but this level was elevated by coinfection with adenovirus particles or by cotransfection with a plasmid which expressed the adenovirus early region 1A (E1A) product. The E1A product is a transcriptional activator and is expressed in 293 cells. Thus, expression from two AAV promoters is differentially regulated: expression from p19 is increased by E1A, w…
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“AAV comprises of a nonenveloped icosahedral viral capsid of ~ 26 nm in diameter, which encapsulates a linear single-stranded DNA genome of ~ 4.7 kb [4].”
From Designing and optimizing AAV-mediated gene therapy for neurodegenerative diseases: from bench to bedside (2024) · cites this paper as [4]
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Metadata
- Authors
- Tratschin, J. D., West, M. H., Sandbank, T., & Carter, B. J.
- Affiliations
- National Institutes of Health (US)
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- Molecular and Cellular Biology
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- Informa UK Limited
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- 1984-10
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- 4
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- 10
- Pages
- 2072-2081
- ISSN
- 0270-7306 (ISSN-L)
- 1098-5549
- PMC
- PMC369024
- OpenAlex
- W1824389368
- Cited by
- 202
- References
- 63
- Citations by year
- 2026: 2 · 2025: 5 · 2024: 7 · 2023: 5 · 2022: 3 · 2021: 6 · 2020: 14 · 2019: 4 · 2018: 4 · 2017: 6 · 2016: 3 · 2015: 2 · 2014: 6 · 2013: 3 · 2012: 3
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- en
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- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Health Sciences › Medicine › Infectious Diseases › Parvovirus B19 Infection Studies
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Keywords
- adeno-associated virus, parvovirus, chloramphenicol acetyltransferase, transient expression, viral encapsidation, recombinant adeno-associated virus, 293 cells, HeLa cells
- MeSH terms
- Acetyltransferases, Chimera, DNA, Viral, Gene Expression Regulation, Genetic Vectors, HeLa Cells, Parvoviridae, Plasmids, Recombination, Genetic, Transfection, Chloramphenicol O-Acetyltransferase
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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